HIV protease inhibitors based on amino acid derivatives

ABSTRACT

A compound selected from the group consisting of a compound of formula I                    
     a compound of formula II                    
     and when the compound of formula I and II comprises an amino group pharmaceutically acceptable ammonium salts thereof, wherein R 1 , R 2 , Cx, n, R 3 , R 4 , R 5 , Y are as defined in the specification.

TECHNICAL FIELD OF THE INVENTION

The present invention concerns amino acid derivatives possessingaspartyl protease inhibitory properties, in particular Nε-amino acidsubstituted L-lysine derivatives (and analogs) possessing aspartylprotease inhibitory properties. It describes also the syntheticmethodology used to make these derivatives and their biologicalapplications. In addition, this invention relates to differentpharmaceutical compositions comprising these compounds. The compoundsand the pharmaceutical compositions of this invention have been shown toinhibit the activity of HIV aspartyl protease, an enzyme essential forvirus maturation. The inhibitory property may be advantageously used toprovide compounds with antiviral properties against HIV viruses,including the HIV-1 and HIV-2 viruses.

BACKGROUND OF THE INVENTION

The HIV (human immunodeficiency virus) retrovirus is responsible forcausing the disease known as AIDS (acquired immunodeficiency syndrome).HIV infection is characterized by a period immediately followinginfection, called asymptomatic, which is devoid of clinicalmanifestations in the patient. Progressive HIV-induced destruction ofthe immune system then leads to increased susceptibility toopportunistic infections, which eventually produces a syndrome calledAIDS-related complex (ARC) characterized by symptoms such as persistentgeneralized lymphadenopathy, fever, weight loss, followed itself by fullblown AIDS.

As the first step of its replication cycle, the HIV-1 retrovirusattaches primarily to the CD4 receptor (a 58 kDa transmembrane protein)to gain entry into susceptible cells, through high-affinity interactionsbetween the viral envelope glycoprotein (gp 120) and a specific regionof the CD4 molecule found in CD4 (+) T-helper lymphocytes and othercells carrying the receptor (Lasky L. A. et al., Cell vol. 50, p.975-985 (1987)). The HIV genetic material, in the form of RNA, is thentranscribed into DNA by a viral enzyme carried by the virus calledreverse transcriptase. The viral DNA now called provirus is thentransported into the cell nucleus in the form of a preintegrationcomplex and attached to the cell DNA by another viral enzyme calledintegrase. Following integration, the viral DNA then serves as atemplate for viral gene expression by the host transcription system. Theprimary RNA transcripts made from the provirus are synthesized by thehost cell RNA polymerase II whose activity is modulated by twovirus-encoded proteins, Tat and Rev. The viral proteins are expressedmainly in the form of polyproteins. After the infected cell has producedall the different HIV polyproteins and genetic material, they assembleat the cell membrane and are released from the cell in the form of animmature viral particle. A third viral enzyme known as protease thencleaves the polyproteins to give the mature, infectious viral particle.The polyproteins that are cleaved by the HIV protease are the Gag andGag-Pol precursors, whose cleavage gives rise to structural proteins andviral enzymes.

A number of synthetic antiviral agents have been designed to blockvarious stages in the replication cycle of HIV, although only thosedeveloped against the viral enzymes have reached the market so far. Thelatter include compounds which block viral reverse transcriptase (forexample, didanosine and zidovudine (AZT)), or the viral protease (forexample, ritonavir and indinavir). Although these drugs have improvedsignificantly the survival time and quality of life of AIDS patients,the administration of most of these agents leads to unwanted sideeffects, such as anemia, neurotoxicity and bone marrow suppression.

Anti-protease compounds represent the most recent drugs developed toblock HIV replication. These compounds inhibit the formation ofinfectious virions by interfering with the processing of viralpolyprotein precursors by the viral protease. The antiviral potential ofHIV protease inhibitors was first demonstrated using peptidicinhibitors. Such peptidic compounds, however, are typically large andcomplex molecules that tend to exhibit poor bioavailability and limitedstability in the body. New compounds devoid of these drawbacks areurgently needed to treat HIV infections. In addition, mutations arisingduring HIV replication lead to resistance to the currently availableprotease inhibitors, so new compounds with original structure aredesirable to fight these resistant viral strains.

SUMMARY OF THE INVENTION

The present invention provides a novel class of compounds, includingtheir pharmaceutically acceptable derivatives. These compounds have anaffinity for aspartyl proteases, in particular, HIV aspartyl protease.Therefore, these compounds are useful as inhibitors of such proteases.These compounds can be used alone or in combination with othertherapeutic or prophylactic agents for the treatment or prophylaxis ofviral infection.

According to a preferred embodiment, the compounds of this invention arecapable of inhibiting HIV viral replication in human CD4+ T-cells, byinhibiting the ability of HIV aspartyl protease to catalyse thehydrolysis of peptide bonds present in viral Gag and Gag-Polpolyproteins. These novel compounds can thus serve to reduce theproduction of infectious virions from acutely and chronically infectedcells, and can inhibit the initial or further infection of host cells.Accordingly, these compounds are useful as therapeutic and prophylacticagents to treat or prevent infection by HIV-1 and HIV-2, which mayresult in asymptomatic infection, AIDS-related complex (ARC), acquiredimmunodeficiency syndrome (AIDS), AIDS-related dementia, or similardiseases of the immune system, and related viruses such as HTLV-I andHTLV-II, and simian immunodeficiency virus.

It is the main objective of this invention to provide a novel class ofmolecules that are aspartyl protease inhibitors, and particularly, HIVaspartyl protease inhibitors.

The present invention relates to a class of Nε-amino acid substitutedL-lysine derivatives (including its lower and higher homologues andanalogs) as well as their pharmaceutically acceptable derivatives (e.g.salts).

Accordingly, the present invention in accordance with one aspect thereofprovides a compound of formula I

a compound of formula II

and when the compound of formula I and II comprises an amino grouppharmaceutically acceptable ammonium salts thereof,

wherein n is 3 or 4

wherein Y is, O, S or N—CN

wherein Cx is selected from the group consisting of —COOM, COOR6, —CHO,—CH₂OR₇, —CH₂OCOR₈, —CONHR9 and —CONR₁₀,R₁₁, wherein M is an alkalimetal (e.g. Na, K, Cs, etc) or alkaline earth metal (Ca, Mg, etc.),

wherein R₁ is selected from the group consisting of a straight orbranched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl grouphaving 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3carbon atoms in the alkyl part thereof,

wherein R₂ is selected from the group consisting of a benzenesulfonylgroup of formula III,

 and a thiophenesulfonyl group of formula IV,

wherein R₃ is selected from the group consisting of H, a straight orbranched alkyl group of 1 to 6 carbon atoms, a phenyl or a benzyl group

wherein R₄ is selected from the group consisting of H, a group offormula IIIa

 a group of formula IVa

 C₆H₁₁—, C₅H₁₀N—CH₂CH₂—, OC₄H₈N—CH₂CH₂— (i.e. morpholine-4-CH₂CH₂—),C₆H₅CH₂CH₂ —, 2,3-(CH ₃O)₂C₆H₃CH₂—, C₆H₅—, 2-C₅H₄N (i.e. 2-pyridinyl),3-C₅H₄N (i.e. 3-pyridinyl), 4-C₅H₄N (i.e. 4-pyridinyl), 3-quinolyl,C₆H₅CS—, 2-naphthyl-SO₂— and a group of formula R_(4C)—CO—, R_(4C) beingselected from the group consisting of a straight or branched alkyl groupof 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbonatoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in thealkyl part thereof, (e.g. CH₃—, iso-butyl, iso-propyl, tert-butyl,tert-butyl-CH₂—), CF₃, 1-pyrrolidinyl, 4-morpholinyl,tetrahydro-3-furanyloxy, 4-CH₃OC₆H₄NH—, CH₃NH—, HOCH₂CH₂NH—,9-fluorenyl-CH₂O—, tert-butylO-, iso-butylO-, C₆H₅CH₂O—, CH₃O—,unsubstituted C₆H₅—, C₆H₅— substituted by one or more members (e.g. oneor two) selected from the group consisting of F, Cl, Br, I, —CF₃, —NO₂,—NR₁₀R₁₁, —NHCOR₁₀, —OR₁₀, —SR₁₀, —COOR₁₀, —COR₁₀ and —CH₂OH,unsubstituted C₆H₅CH₂—, C₆H₅CH₂— substituted by one or more members(e.g. one or two) selected from the group consisting of F, Cl, Br, I,—CF₃, —NO₂, —NR₁₀R₁₁, —NHCOR₁₀, —OR₁₀, —SR₁₀, —COOR₁₀, —COR₁₀ and—CH₂OH, unsubstituted C₆H₅CH₂CH₂—, and C₆H₅CH₂CH₂— substituted by one ormore members (e.g. one or two) selected from the group consisting of F,Cl, Br, I, —CF₃, —NO₂, —NR₁₀R₁₁, —NHCOR₁₀, —OR₁₀, —SR₁₀, —COOR₁₀, —COR₁₀and —CH₂OH,

wherein R₅ is selected from the group consisting of H, a straight orbranched alkyl group of 1 to 8 carbon atoms, (e.g. CH₃—, CH₃CH₂CH₂—,CH₃CH₂CH₂CH₂—, iso-C₄H₉—, C₆H₁₁CH₂—), HOCH₂—, C₆H₅CH₂OCH₂—,benzyl-OCH(CH₃), HO₂CCH₂—, HO₂CCH₂CH₂—, NC—CH₂—, H₂NC(O)CH₂—,H₂NC(O)CH₂CH₂—, 4-CH₃C₆H₄CH₂SCH₂—, CH₃SCH₂CH₂—, H₂NCH₂CH₂CH₂CH₂—, C₆H₅—,C₆H₅CH₂—, C₆H₅CH(OH)—, C₆H₅CH(CN)—, C₆F₅CH₂—,4-(9-fluorenylmethoxycarbonyl)-NHCH₂—C₆H₄CH₂—, C₅H₄N-2-CH₂— (i.e.pyridine-2-CH₂—), C₅H₄N-3-CH₂— (i.e. pyridine-3-CH₂—), C₅H₄N-4-CH₂—(i.e. pyridine-4-CH₂—), 2-thiophene-CH₂—, indole-3-CH₂—,2-benzothiophene-CH₂—, Nτ-benzyl-imidazole-4-CH₂—, imidazole-4-CH₂—,thiazole-4-CH₂— and substituted benzyl (e.g. benzyl substituted by agroup being as defined for R₁₂ below, e.g. 4-tert-butyl-C₆H₄CH₂—,4-HOC₆H₄CH₂—, 4-benzyl-O—C₆H₄CH₂—, 4-NO₂C₆H₄CH₂—, 2-FC₆H₄CH₂—,3-FC₆H₄CH₂—, 4-FC₆H₄CH₂—),

wherein Ra represents a member selected from the group consisting of

 wherein Met is a methylene linked to the α′ nitrogen

wherein R₆ is selected from the group consisting of H, a straight orbranched alkyl group of 1 to 4 carbon atoms and glycyl

wherein R₇ is selected from the group consisting of H, a straight orbranched alkyl group of 1 to 4 carbon atoms

wherein R₈ is selected from the group consisting of a straight orbranched alkyl group of 1 to 6 carbon atoms, a cycloalkyl group having 3to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms inthe cycloalkyl part and 1 to 3 carbon atoms in the alkyl part thereof,

wherein R₉ is selected from the group consisting of H, a straight orbranched alkyl group of 1 to 4 carbon atoms, —OH, —NH₂ and —CH₂CH₂OH

wherein R₁₀ and R₁₁ are independently selected from the group consistingof H, a straight or branched alkyl group of 1 to 4 carbon atoms

wherein m is 0 or 1

wherein o is 0 or 1

wherein R₁₂ and R₁₃ are independently selected from the group consistingof H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl,Br, I, —CF₃, —NO₂, —NR₁₀ R₁₁, —NHCOR₁₀, —OR₁₀, —OCH₂C₆H₅, —SR₁₀,—COOR₁₀, —COR₁₀and —CH₂OH, R₁₀ and R₁₁ being as defined herein.

More particularly, this invention provides a compound of formula IA

and when the compound of formula IA comprises an amino grouppharmaceutically acceptable ammonium salts thereof,

wherein n is 3 or 4

wherein Y is O, S or N—CN

wherein Cx is selected from the group consisting of —COOM, COOR₆, —CHO,—CH₂OR₇, —CH₂OCOR₈, —CONHR₉ and —CONR₁₀R₁₁, wherein M is an alkali metal(e.g. Na, K, Cs, etc) or alkaline earth metal,

wherein R₁ is selected from the group consisting of a straight orbranched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl grouphaving 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3carbon atoms in the alkyl part thereof,

wherein R₂ is selected from the group consisting of a benzenesulfonylgroup of formula III,

 and a thiophenesulfonyl group of formula IV,

wherein R₃ is selected from the group consisting of H, a straight orbranched alkyl group of 1 to 6 carbon atoms, a phenyl or a benzyl group

wherein R₄ is selected from the group consisting of H, C₆H₁₁—,C₅H₁₀N—CH₂CH₂—, OC₄H₈N—CH₂CH₂— (i.e. morpholine-4-CH₂CH₂—), C₆H₅CH₂CH₂—,2,3-(CH₃O)₂C₆H₃CH₂—, C₆ _(H) ₅—, 2-C₅H₄N, 3-C₅H₄N, 4-C₅H₄N, 3-quinolyl,CH₃CO—, CF₃CO, C₆H₅CO—, C₆H₅CS—, 4CH₃OC₆H₄CH₂CO—, C₆H₅CH₂CH₂CO—,iso-butyl-CO—, iso-propyl-CO—, tert-butyl-CO—, tert-butyl-CH₂CO—,1-pyrrolidine-CO—, 4-morpholine-CO—, carbotetrahydro-3-furanyloxy,4-CH₃OC₆H₄NHCO—, CH₃NHCO—, HOCH₂CH₂NHCO—, 9-fluorenylmethoxycarbonyl,tert-butylO-CO—, iso-butylO-CO—, C₆H₅CH₂O—CO—, CH₃O—CO—, C₆H₅SO₂—,4-CH₃C₆H₄SO₂—, 4-CF₃C₆H₄SO₂—, 4-NO₂C₆H₄SO₂—, 4-NH₂C₆H₄SO₂—,4-AcNHC₆H₄SO₂—, 4-FC₆H₄SO₂—, 4-ClC₆H₄SO₂—, 4-BrC₆H₄SO₂—, 4-CH₃OC₆H₄SO₂—,2-thiophene-SO₂— and 2-naphthyl-SO₂—

wherein R₅ is selected from the group consisting of H, CH₃—, CH₃CH₂CH₂—,CH₃CH₂CH₂CH₂—, iso-C₄H₉—, C₆H₁₁CH₂—, HOCH₂—, C₆H₅CH₂OCH₂—,benzyl-OCH(CH₃), HO₂CCH₂—, HO₂CCH₂CH₂—, NC—CH₂—, H₂NC(O)CH₂—,H₂NC(O)CH₂CH₂—, 4-CH₃C₆H₄CH₂SCH₂—, CH₃SCH₂CH₂—_(,) H₂NCH₂CH₂CH₂CH₂—,C₆H₅—, C₆H₅CH₂—, C₆F₅CH₂—, 4-tert-butyl-C₆H₄CH₂—, 4-HOC₆H₄CH₂—,4-benzyl-O—C₆H₄CH₂—, 4-NO₂C₆H₄CH₂—,4-(9-fluorenylmethoxycarbonyl)NHCH₂—C₆H₄CH₂—, 2-FC₆H₄CH₂—, 3-FC₆H₄CH₂—,4-FC₆H₄CH₂—, C₅H₄N-2-CH₂—, C₅H₄N-3-CH₂—, C₅H₄N-4-CH₂—,2-thiophene-CH₂—_(,) indole-3-CH₂—, 2-benzothiophene-CH₂—,Nτ-benzyl-imidazole-4-CH₂— and thiazole-4-CH₂—

wherein R₆ is selected from the group consisting of H, a straight orbranched alkyl group of 1 to 4 carbon atoms and glycyl

wherein R₇ is selected from the group consisting of H, a straight orbranched alkyl group of 1 to 4 carbon atoms

wherein R₈ is selected from the group consisting of a straight orbranched alkyl group of 1 to 6 carbon atoms, a cycloalkyl group having 3to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms inthe cycloalkyl part and 1 to 3 carbon atoms in the alkyl part thereof,

wherein R₉ is selected from the group consisting of H, a straight orbranched alkyl group of 1 to 4 carbon atoms, —OH, —NH₂ and —CH₂CH₂OH

wherein R₁₀ and R₁₁ are independently selected from the group consistingof H, a straight or branched alkyl group of 1 to 4 carbon atoms

wherein m is 0 or 1

wherein o is 0 or 1

wherein R₁₂ and R₁₃ are independently selected from the group consistingof H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl,Br, I, —CF₃, —NO₂, —NR₁₀R₁₁, —NHCOR₁₀, —OR₁₀, —OCH₂C₆H₅, —SR₁₀, —COOR₁₀,—COR₁₀ and —CH₂OH, R₁₀ and R₁₁ being as defined herein.

This invention also provides a compound of formula Ia

and when the compound of formula Ia comprises an amino grouppharmaceutically acceptable ammonium salts thereof,

wherein Cx is selected from the group consisting of —COOM and —CH₂OH, Mbeing an alkali metal or alkaline earth metal,

wherein R_(1A) is selected from the group consisting of a straight orbranched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl grouphaving 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3carbon atoms in the alkyl part thereof,

wherein R_(2A) and R_(4A) are independently selected from the groupconsisting of H, a straight or branched alkyl group of 1 to 4 carbonatoms, F, Cl, Br, I, —CF₃, —NO₂, —NR₁₀R₁₁, —NHCOR₁₀, —OR₁₀, —OCH₂C₆H₅,—SR₁₀, —COOR₁₀, —COR₁₀ and —CH₂OH,

wherein R_(5A) is selected from the group consisting of H, a straight orbranched alkyl group of 1 to 8 carbon atoms, (e.g. CH₃—, CH₃CH₂CH₂—,CH₃CH₂CH₂CH₂—, iso-C₄H₉—, C₆H₁₁CH₂—, HOCH₂—), C₆H₅CH₂OCH₂—,benzyl-OCH(CH₃), HO₂CCH₂—, HO₂CCH₂CH₂—, NC—CH₂—, H₂NC(O)CH₂—,H₂NC(O)CH₂CH₂—, 4-CH₃C₆H₄CH₂SCH₂—, CH₃SCH₂CH₂—_(,) H₂NCH₂CH₂CH₂CH₂—,C₆H₅—, C₆H₅CH₂—, C₆H₅CH(OH)—, C₆H₅CH(CN)—, C₆F₅CH₂—,4-(9-fluorenylmethoxycarbonyl)-NHCH₂—C₆H₄CH₂—, C₅H₄N-2-CH₂— (i.e.pyridine-2-CH₂—), C₅H₄N-3-CH₂— (i.e. pyridine-3-CH₂—), C₅H₄N-4-CH₂—(i.e. pyridine-4-CH₂—), 2-thiophene-CH₂—_(,) indole-3-CH₂—,2-benzothiophene-CH₂—, Nτ-benzyl-imidazole-4-CH₂—, imidazole-4-CH₂—,thiazole-4-CH₂— and substituted benzyl (e.g. benzyl substituted by agroup as defined for R_(2A), e.g. 4-tert-butyl-C₆H₄CH₂—, 4-HOC₆H₄CH₂—,4-benzyl-O-C₆H₄CH₂—, 4-NO₂C₆H₄CH₂—, 2-FC₆H₄CH₂—, 3-FC₆H₄CH₂—,4-FC₆H₄CH₂—),

and wherein R₁₀ and R₁₁ are as defined herein.

This invention also provides a compound of formula Ib

and when the compound of formula Ib comprises an amino grouppharmaceutically acceptable ammonium salts thereof,

wherein Cx is selected from the group consisting of —COOM, and —CH₂OH, Mbeing an alkali metal or alkaline earth metal,

wherein R_(1A) is selected from the group consisting of a straight orbranched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl grouphaving 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3carbon atoms in the alkyl part thereof,

wherein R_(2A) and R_(4B) are independently selected from the groupconsisting of H, a straight or branched alkyl group of 1 to 4 carbonatoms, F, Cl, Br, I, —CF₃, —NO₂, —NR₁₀R₁₁, —NHCOR₁₀, —OR₁₀, —OCH₂C₆H₅,—SR₁₀, —COOR₁₀, —COR₁₀ and —CH₂OH,

wherein R_(5A) is selected from the group consisting of H, a straight orbranched alkyl group of 1 to 8 carbon atoms, (e.g. CH₃—, CH₃CH₂CH₂—,CH₃CH₂CH₂CH₂—, iso-C₄H₉—, C₆H₁₁CH₂—), HOCH₂—, C₆H₅CH₂OCH₂—,benzyl-OCH(CH₃), HO₂CCH₂—, HO₂CCH₂CH₂—, NC—CH₂—, H₂NC(O)CH₂—,H₂NC(O)CH₂CH₂—, 4-CH₃C₆H₄CH₂SCH₂—, CH₃SCH₂CH₂—, H₂NCH₂CH₂CH₂CH₂—, C₆H₅—,C₆H₅CH₂—, C₆H₅CH(OH)—, C₆H₅CH(CN)—, C₆F₅CH₂—,4-(9-fluorenylmethoxycarbonyl)-NHCH₂—, C₆H₄CH₂—, C₅H₄N-2-CH₂— (i.e.pyridine-2-CH₂—), C₅H₄N-3-CH₂— (i.e. pyridine-3-CH₂—), C₅H₄N-4-CH₂—(i.e. pyridine-4-CH₂—), 2-thiophene-CH₂—_(,) indole-3-CH₂—,2-benzothiophene-CH₂—, Nτ-benzyl-imidazole-4-CH₂—, imidazole-4-CH₂—,thiazole-4-CH₂— and substituted benzyl (e.g. benzyl substituted by agroup as defined for R_(2A), e.g. 4-tert-butyl-C₆H₄CH₂—, 4-HOC₆H₄CH₂—,4-benzyl-O—C₆H₄CH₂—, 4-NO₂C₆H₄CH₂—, 2-FC₆H₄CH₂—, 3-FC₆H₄CH₂—,4-FC₆H₄CH₂—),

and wherein R₁₀ and R₁₁ are as defined herein.

This invention also provides a compound of formula Ic

and when the compound of formula Ic comprises an amino grouppharmaceutically acceptable ammonium salts thereof,

wherein Cx is selected from the group consisting of —COOM and —CH₂OH, Mbeing an alkali metal or alkaline earth metal,

wherein R_(1A) is selected from the group consisting of a straight orbranched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl grouphaving 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3carbon atoms in the alkyl part thereof,

wherein R_(2B) and R_(4B) are independently selected from the groupconsisting of H, a straight or branched alkyl group of 1 to 4 carbonatoms, F, Cl, Br, I, —CF₃, —NO₂, —NR₁₀R₁₁, —NHCOR₁₀, —OR₁₀, —OCH₂C₆H₅,—SR₁₀, —COOR₁₀, —COR₁₀ and —CH₂OH,

wherein R_(5A) is selected from the group consisting of H, a straight orbranched alkyl group of 1 to 8 carbon atoms, (e.g. CH₃—, CH₃CH₂CH₂—,CH₃CH₂CH₂CH₂—, iso-C₄H₉—, C₆H₁₁CH₂—), HOCH₂—, C₆H₅CH₂OCH₂—,benzyl-OCH(CH₃), HO₂CCH₂—, HO₂CCH₂CH₂—, NC—CH₂—, H₂NC(O)CH₂—,H₂NC(O)CH₂CH₂—, 4-CH₃C₆H₄CH₂SCH₂—, CH₃SCH₂CH₂—_(,) H₂NCH₂CH₂CH₂CH₂—,C₆H₅—, C₆H₅CH₂—, C₆H₅CH(OH)—, C₆H₅CH(CN)—, C₆F₅CH₂—,4-(9-fluorenylmethoxycarbonyl)-NHCH₂—C₆H₄CH₂—, C₅H₄N-2-CH₂— (i.e.pyridine-2-CH₂—), C₅H₄N-3-CH₂— (i.e. pyridine-3-CH₂—), C₅H₄N-4-CH₂—(i.e. pyridine-4-CH₂—), 2-thiophene-CH₂—_(,) indole-3-CH₂—,2-benzothiophene-CH₂—, Nτ-benzyl-imidazole-4-CH₂—, imidazole-4-CH₂—,thiazole-4-CH₂— and substituted benzyl (e.g. benzyl substituted by agroup as defined for R_(2A), e.g. 4-tert-butyl-C₆H₄CH₂—, 4-HOC₆H₄CH₂—,4-benzyl-O—C₆H₄CH₂—, 4-NO₂C₆H₄CH₂—, 2-FC₆H₄CH₂—, 3-FC₆H₄CH₂ —, 4-FC₆H₄CH₂—),

and wherein R₁₀ and R₁₁ are as defined herein.

This invention also provides a compound of formula Id

and when the compound of formula Id comprises an amino grouppharmaceutically acceptable ammonium salts thereof,

wherein Cx is selected from the group consisting of —COOM and —CH₂OH, Mbeing an alkali metal or alkaline earth metal,

wherein R_(1A) is selected from the group consisting of a straight orbranched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl grouphaving 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3carbon atoms in the alkyl part thereof,

wherein R_(4A) is selected from the group consisting of H, a straight orbranched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, —CF₃, —NO₂,—NR₁₀R₁₁, —NHCOR₁₀, —OR₁₀, —OCH₂C₆H₅, —SR₁₀, —COOR₁₀, —COR₁₀ and —CH₂OH,

wherein R_(4C) is selected from the group consisting of a straight orbranched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl grouphaving 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3carbon atoms in the alkyl part thereof, (e.g. CH₃—, iso-butyl,iso-propyl, tert-butyl, tert-butyl-CH₂—), CF₃, pyrrolidine,4-morpholine, tetrahydro-3-furanyloxy, 4-CH₃OC₆H₄NH—, CH₃NH—,HOCH₂CH₂NH—, 9-fluorenyl-CH₂O—, tert-butylO-, iso-butylO-, C₆H₅CH₂O—,CH₃O—, unsubstituted C₆H₅—, C₆H₅— substituted by one or more members(e.g. one or two) selected from the group consisting of F, Cl, Br, I,—CF₃, —NO₂, —NR₁₀R₁₁, —NHCOR₁₀, —OR₁₀, —SR₁₀, —COOR₁₀, —COR₁₀ and—CH₂OH, unsubstituted C₆H₅CH₂—, C₆H₅CH₂— substituted by one or moremembers (e.g. one or two) selected from the group consisting of F, Cl,Br, I, —CF₃, —NO₂, —NR₁₀R₁₁, —NHCOR₁₀, —OR₁₀, —SR₁₀, —COOR₁₀, —COR₁₀ and—CH₂OH, unsubstituted C₆H₅CH₂CH₂—, and C₆H₅CH₂CH₂— substituted by one ormore members (e.g. one or two) selected from the group consisting of F,Cl, Br, I, —CF₃, —NO₂, —NR₁₀R_(11, —NHCOR) ₁₀, —OR₁₀, —SR₁₀, —COOR₁₀,—COR₁₀ and —CH₂OH,

wherein R_(5A) is selected from the group consisting of H, a straight orbranched alkyl group of 1 to 8 carbon atoms, (e.g. CH₃—, CH₃CH₂CH₂—,CH₃CH₂CH₂CH₂—, iso-C₄H₉—, C₆H₁₁CH₂—), HOCH₂—, C₆H₅CH₂OCH₂—,benzyl-OCH(CH₃), HO₂CCH₂—, HO₂CCH₂CH₂—, NC—CH₂—, H₂NC(O)CH₂—,H₂NC(O)CH₂CH₂—, 4-CH₃C₆H₄CH₂SCH₂—, CH₃SCH₂CH₂—_(,) H₂NCH₂CH₂CH₂CH₂—,C₆H₅—, C₆H₅CH₂—, C₆H₅CH(OH)—, C₆H₅CH(CN)—, C₆F₅CH₂—,4-(9-fluorenylmethoxycarbonyl)-NHCH₂—C₆H₄CH₂—, C₅H₄N-2-CH₂— (i.e.pyridine-2-CH₂—), C₅H₄N-3-CH₂— (i.e. pyridine-3-CH₂—), C₅H₄N-4-CH₂—(i.e. pyridine-4-CH₂—), 2-thiophene-CH₂—_(,) indole-3-CH₂—,2-benzothiophene-CH₂—, Nτ-benzyl-imidazole-4-CH₂—, imidazole-4-CH₂—,thiazole-4-CH₂— and substituted benzyl (e.g. benzyl substituted by agroup as defined for R_(2A), e.g. 4-tert-butyl-C₆H₄CH₂—, 4-HOC₆H₄CH₂—,4-benzyl-O—C₆H₄CH₂—, 4-NO₂C₆H₄CH₂—, 2-FC₆H₄CH₂—, 3-FC₆H₄CH₂—,4-FC₆H₄CH₂—),

and wherein R₁₀ and R₁₁ are as defined herein.

In addition, this invention provides pharmaceutical compositions inwhich these novel compounds of formula I or II, (as well as of formulaIA, Ia, Ib, Ic and Id) derived from L-lysine or L-lysine derivatives (aswell as lower and higher homologues) are used to inhibit aspartylproteases, including HIV aspartyl protease, thus providing protectionagainst HIV infection.

Thus the present invention provides a pharmaceutical compositioncomprising a pharmaceutically acceptable carrier and a pharmaceuticallyeffective amount of at least one compound of formula I, formula II (aswell as of formulae IA, Ia, Ib, Ic, and Id) and as applicablepharmaceutically acceptable ammonium salts thereof.

The terms “HIV protease” and “HIV aspartyl protease” are usedinterchangeably and refer to the aspartyl protease encoded by the humanimmunodeficiency virus type 1 or 2. In a preferred embodiment of thisinvention, these terms refer to the human immunodeficiency virus type 1aspartyl protease.

The term “pharmaceutically effective amount” refers to an amounteffective in treating HIV infection in a patient.

The term “prophylactically effective amount” refers to an amounteffective in preventing HIV infection in a patient. As used herein, theterm “patient” refers to a mammal, including a human.

The terms “pharmaceutically acceptable carrier”, “pharmaceuticallyacceptable adjuvant” and “physiologically acceptable vehicle” refer to anon-toxic carrier or adjuvant that may be administered to a patient,together with a compound of this invention, and which does not destroythe pharmacological activity thereof.

The compounds of this invention include pharmaceutically acceptablederivatives of the compounds of formula I, formula II (as well as offormulae IA, Ia, Ib, Ic, and Id) and as applicable pharmaceuticallyacceptable ammonium salts thereof. A “pharmaceutically acceptablederivative” means any pharmaceutically acceptable salt, ester, or saltof such ester, of a compound of this invention or any other compoundwhich, upon administration to a recipient, is capable of providing(directly or indirectly) a compound of this invention or an antivirallyactive metabolite or residue thereof.

Salts derived from appropriate bases include alkali metal (e.g.,sodium), alkaline earth metal (e.g., magnesium), ammonium and N—(C₁₋₄alkyl)₄ ⁺ salts.

The compounds of this invention contain one or more asymmetric carbonatoms and thus may occur as racemates and racemic mixtures, singleenantiomer, diastereomeric mixtures and individual diastereoisomers. Allsuch isomeric forms of these compounds are expressly included in thepresent invention. Each stereogenic carbon may be of the R or Sconfiguration.

Combinations of substituents and variables envisioned by this inventionare only those that result in the formation of stable compounds. Theterm “stable”, as used herein, refers to compounds which possessstability sufficient to allow manufacture and administration to a mammalby methods known in the art. Typically, such compounds are stable at atemperature of 40° C. or less, in the absence of moisture or otherchemically reactive conditions, for at least a week.

Pharmaceutically acceptable salts of the compounds of this inventioninclude those derived from pharmaceutically acceptable inorganic andorganic acids and bases. Examples of such acid salts include: acetate,adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate,butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate,digluconate, dodecylhydrogensulfate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptanoate, glycerophosphate, glycollate,hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide,hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, malonate,methanesulfonate, 2-naphthylsulfonate, nicotinate, nitrate, oxalate,pamoate, pectinate, perchlorate, persulfate, 3-phenylpropionate,phosphate, picrate, pivalate, propionate, salicylate, succinate,sulfate, tartrate, thiocyanate, tosylate, and undecanoate.

This invention also envisions the quaternization of any basic nitrogencontaining groups of the compounds disclosed herein. The basic nitrogencan be quaternized with any agents known to those of ordinary skill inthe art including, for example, lower alkyl halides, such as methyl,ethyl, propyl and butyl chlorides, bromides and iodides; dialkylsulfates including dimethyl, diethyl, dibutyl and diamyl sulfates; longchain halides such as decyl, lauryl, myristyl and stearyl chlorides,bromides and iodides, and aralkyl halides including benzyl and phenethylbromides. Water or oil-soluble or dispersible products may be obtainedby such quaternization.

The compounds of this invention are readily prepared using conventionaltechniques from commercially available and inexpensive startingmaterials. The relative ease of synthesis of the products describedherein represents a marked advantage for the large scale preparation ofthese compounds. In general, the derivatives of the present inventionmay be readily obtained through sequences recognized by thoseknowledgeable in the art as straightforward. These sequences arepresented in schemes 1 to 7 discussed below.

Scheme 1 illustrates a generic example for the preparation of a keyintermediate needed for the synthesis of HIV protease inhibitors.

Note:

a) For scheme 1, R₁ represents an alkyl or cycloalkylalkyl side chain asdefined above

b) R₂ represents a benzenesulfonyl group of formula III, athiophenesulfonyl group of formula IV, a 1-naphthylsulfonyl, a2-naphthylsulfonyl or a 8-quinolinesulfonyl group as defined above

As shown in scheme 1, the Nα,Nα-disubstituted L-lysine derivative 5 wasobtained from commercially available L-lysine 1 in a four-step reactionsequence. This preparation uses the cyclic form of L-lysine in order tomanipulate the Nα-amino group without the need for protective groups.First, L-lysine was transformed into L-α-amino-ε-caprolactam 2 upontreatment with hydrochloric acid in methanol followed by neutralizationwith sodium hydroxide. The caprolactam 2 is also commercially available.Reductive alkylation of derivative 2 with an appropriate aldehyde andNaBH(OAc)₃ in dichloroethane lead to the Nα-alkylamino-ε-caprolactam 3.Then, sulfonation with an arylsulfonyl chloride or asubstituted-arylsulfonyl chloride in the presence of triethylamine indichloromethane gave compound 4 in excellent yields. TheNα,Nα-disubstituted L-lysine derivative 5 was obtained quantitatively byacid hydrolysis of the cyclic amide 4.

Scheme 2 illustrates the preparation of HIV protease inhibitors bearingeither a carboxylic function, compound 6, or an alcohol function,compound 8, on the final product. In other words, this scheme shows thesynthesis of a L-lysine derivative or a (2S) 2,6-diaminohexanolderivative

Note:

a) For scheme 2, R₁ and R₂ are as defined above

b) R₃ represents H, a straight or branched alkyl group of 1 to 6 carbonatoms, a phenyl or a benzyl group

c) R₄ is as defined above

d) R₅ represents an amino acid side chain as defined above

Following the indications summarized in Scheme 2, derivative 5 is linkedwith a substituted amino acid using N,N-carbonyldiimidazole as theactivating reagent to yield derivative 6 in good to excellent yields.The various N-acylated (or N-sulfonated) amino acids needed for thecoupling reaction were prepared from the appropriate amino acid and acylchloride (or sulfonyl chloride) using the Schotten-Baumen procedure.Alternatively, derivative 5 is treated with trimethylsilyl chloride inmethanol (HCl generated in situ) and the resulting methyl esterintermediate is reduced with lithium aluminum hydride (LAH) in THF toafford 7 in good yields. The (2S) 2,6-diaminohexanol derivative 7 islinked to a substituted amino acid derivative as it is described abovefor the synthesis of derivative 6.

The L-lysine divative 6 can be further transformed into a variety ofesters 9 as well as amide derivative 10 as shown in scheme 3. Thesetransformations are done under standard reaction conditions. Forexample, the synthesis of ester 9 can be achieved upon activation of theacid 6 with DCC in the presence of a catalytic amount ofN,N-dimethylaminopyridine and an alcohol. The amide 10 can be obtainedas described earlier for the preparation of compound 6, see scheme 2.

Note:

a) For scheme 3, R₁, R₂, R₃, R₄ and R₅ are as described above

b) R₁₀ and R₁₁, same or different, represent an H or a straight orbranched alkyl group of 1 to 4 carbon atoms.

Scheme 4 presents a second approach for the preparation of HIV proteaseinhibitors of formula 6 and 8. It proceeds by using commerciallyavailable Ne-benzyloxycarbonyl-L-lysine methyl ester hydrochloride (11)as the starting material. Reductive alkylation of derivative 11 with anappropriate aldehyde and sodium cyanoborohydride provided the derivative12. Then, sulfonation with benzenesulfonyl chloride (orsubstituted-benzenesulfonyl chloride) in the presence of triethylamine(or diisopropylethylamine) in dichloromethane gave compound 13 inexcellent yields for the two first steps. Removal of thebenzyloxycarbonyl group (Z group) by hydrogen gas in presence of 10%Pd/C yielded the free Nε-amino derivative 14 quantitatively. Acylationof 14 with a substituted amino acid N-hydroxysuccinimide ester providedderivative 15 in excellent yields. The desired HIV protease inhibitors 6and 8 are easily obtained from the methyl ester 15 by hydrolysis withsodium hydroxide in a mixture of THF and methanol giving the acid 6 orby reduction with LAH giving the alcohol 8, both in excellent yields. Itis noteworthy that, under basic hydrolysis of 15 to produce compound 6,some racemization may occur. However, it is not the case when compound15 is reduced with LAH to give derivative 8.

Note:

a) For scheme 4, R₁, R₂, R₃, R₄ and R₅ are as described above

Scheme 5 illustrates the preparation of an anti-protease derivativeusing a solid phase methodology in accordance with the present invention(see example 21). Any suitable solid phase substrate could be used insuch preparation (K. Burgess, Solid phase organic synthesis,Wiley-Interscience, 2000).

Note:

a) For scheme 4, R₁ is iso-butyl, R₂ is 4-methylbenzenesulfonyl, R₃, R₄and R₅ are as described above

This process allows the introduction of pharmacophores to aNα,Nα-disubstituted-L-lysine derivative (such as 16) via the N-terminalfunction. Thus, in scheme 5,Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysine16 is immobilized on a p-benzyloxybenzylalcohol resin (Wang resin) inDMF for a period of 16 h. The resulting component 17 contained 0.28 meq.of L-lysine derivative/g of resin. At this stage, after removal of theFmoc protective group under standard reaction conditions (30% piperidinein DMF see T. W. Greene and P. G. M. Wuts, Protective groups in OrganicSynthesis, 3^(rd) Edition, John Wiley & Sons, Inc. 2000), the resin canbe coupled with a variety of N-acylated (or N-sulfonated) amino acids togive component 18. The N-acylated (or N-sulfonated) amino acids areactivated with N-hydroxysuccinimide and DCC in DMF. Cleavage of theresin with TFA in CH₂Cl₂ leads to the desired L-lysine derivative 19.

Scheme 6 illustrates the preparation of substituted glycine derivativesused for the synthesis of several HIV protease inhibitors in accordancewith the present invention (see examples 114 and 158 below for specificdescriptions of the synthesis of such glycine derivatives):

In scheme 6a), N-phenylglycine 20 is treated with an excess butyllithiumto give the dianion intermediate to which an appropriate alkyl halide(or arylalkyl halide or tosylate) is added and reacted for a period of16 h. The final products 21 are obtained in good to excellent yields. Anappropriate alkyl halide is defined as bearing a R₃ component which cansustain strong basic reaction conditions.

In scheme 6b), methyl bromoacetate 22 is treated with benzylamine inCH₂Cl₂ at room temperature for 16 h. The N-benzylglycine methyl esterderivative 23 was obtained in 86% yield. This intermediate can be eitheracylated with a carboxylic acid derivative and DCC in THF or sulfonatedwith an appropriate sulfonyl chloride and triethylamine in CH₂Cl₂ togive derivative 24 or 25 as desired in good to excellent yields.

Scheme 7 shows another methodology for the formation ofNε-substituted-glycyl-L-lysine HIV protease inhibitors via theNε-iodoacetyl-L-lysine derivative 26 (see example 105 for the detaileddescription of the synthesis of derivative 26 and its use). Thus,Nα,Nα-disubstituted-L-lysine derivative 5 potassium salt is initiallytreated with chloroacetyl chloride in the presence of DIEA in THF togive the Nα,Nα-disubstituted-Nε-chloroacetyl-L-lysine intermediate. Thisintermediate is transformed into the iodoacetyl derivative 26 upontreatment with sodium iodide in dry acetone. Compound 26 is then heatedat reflux with a primary (or secondary) amine in the presence of of DIEAin THF to yield the desired Nε-substituted-glycyl-L-lysine derivative 6.In scheme 7, a primary amine is used so R₃=H and R₅=H.

As it can be appreciated by the skilled artisan, the above syntheticschemes are not intended to be a comprehensive list of all means bywhich the compounds described and claimed in this application may besynthesized. Further methods will be evident to those of ordinary skillin the art.

The compounds of this invention may be modified by appending appropriatefunctionalities to enhance selective biological properties. Suchmodifications are known in the art and include those which increasebiological penetration into a given biological system (e.g., blood,lymphatic system, central nervous system), increase oral availability,increase solubility to allow administration by injection, altermetabolism and alter rate of excretion.

As discussed above, the novel compounds of the present invention areexcellent ligands for aspartyl proteases, particularly HIV-1 protease.Accordingly, these compounds are capable of targeting and inhibitinglate stage events in the replication, i.e. the processing of the viralpolyproteins by HIV encoded protease. Compounds according to thisinvention advantageously inhibit the ability of the HIV-1 virus toinfect immortalized human T cells over a period of days, as determinedby an assay measuring the amount of extracellular p24 antigen—a specificmarker of viral replication (see, Meek et al., Nature, 343, pp. 90-92(1990)).

In addition to their use in the prophylaxis or treatment of HIV or HTLVinfection, the compounds according to this invention may also be used asinhibitory or interruptive agents for other viruses which depend onaspartyl proteases, similar to HIV or HTLV aspartyl proteases, forobligatory events in their life cycle. Such compounds inhibit theproteolytic processing of viral polyprotein precursors by inhibitingaspartyl protease. Because aspartyl protease is essential for theproduction of mature virions, inhibition of that processing effectivelyblocks the spread of virus by inhibiting the production and reproductionof infectious virions, particularly from acutely and chronicallyinfected cells. The compounds of this invention advantageously inhibitaspartyl proteases, thus blocking the ability of aspartyl proteases tocatalyse the hydrolysis of peptide bonds.

The compounds of this invention may be employed in a conventional mannerfor the treatment or prevention of HIV, HTLV, and other viruses, whichdepend on aspartyl proteases for obligatory events in their life cycle.Such methods of treatment, their dosage levels and requirements may beselected by those of ordinary skill in the art from available methodsand techniques. For example, a compound of this invention may becombined with a pharmaceutically acceptable adjuvant for administrationto a virally infected patient in a pharmaceutically acceptable mannerand in an amount effective to lessen the severity of the viralinfection.

Alternatively, the compounds of this invention may be used in vaccinesand methods for protecting individuals against viral infection over anextended period of time. The compounds may be employed in such vaccineseither alone or together with other compounds of this invention in amanner consistent with the conventional utilization of proteaseinhibitors in vaccines. For example, a compound of this invention may becombined with pharmaceutically acceptable adjuvants conventionallyemployed in vaccines and administered in prophylactically effectiveamounts to protect individuals over an extended period of time againstviral infections, such as HIV infection. As such, the novel proteaseinhibitors of this invention can be administered as agents for treatingor preventing viral infections, including HIV infection, in a mammal.

The compounds of this invention may be administered to a healthy orHIV-infected patient either as a single agent or in combination withother antiviral agents which interfere with the replication cycle ofHIV. By administering the compounds of this invention with otherantiviral agents which target different events in the viral life cycle,the therapeutic effect of these compounds is potentiated. For instance,the co-administered antiviral agent can be one which targets earlyevents in the viral life cycle, such as attachment to the cell receptorand cell entry, reverse transcription and viral DNA integration intocellular DNA. Antiviral agents targeting such early life cycle eventsinclude among others polysulfated polysaccharides, sT4 (soluble CD4) andother compounds which block binding of virus to CD4 receptors on CD4bearing T-lymphocytes and other CD4(+) cells, or inhibit fusion of theviral envelope with the cytoplasmic membrane, and didanosine (ddI),zalcitabine (ddC), stavudine (d4T), zidovudine (AZT) and lamivudine(3TC) which inhibit reverse transcription. Other anti-retroviral andantiviral drugs may also be co-administered with the compounds of thisinvention to provide therapeutic treatment for substantially reducing oreliminating viral infectivity and the symptoms associated therewith.Examples of other antiviral agents include ganciclovir, dideoxycytidine,trisodium phosphonoformate, eflornithine, ribavirin, acyclovir, alphainterferon and trimenotrexate. Additionally, other types of drugs may beused to potentiate the effect of the compounds of this invention, suchas viral uncoating inhibitors, inhibitors of Tat or Rev trans-activatingproteins, antisense molecules or inhibitors of the viral integrase.These compounds may also be co-administered with other inhibitors of HIVaspartyl protease.

Combination therapies according to this invention exert a synergisticeffect in inhibiting HIV replication because each component agent of thecombination acts on a different site of HIV replication. The use of suchcombinations also advantageously reduces the dosage of a givenconventional anti-retroviral agent that would be required for a desiredtherapeutic or prophylactic effect as compared to when that agent isadministered as a monotherapy. These combinations may reduce oreliminate the side effects of conventional single anti-retroviral agenttherapies while not interfering with the anti-retroviral activity ofthose agents. These combinations reduce the potential of resistance tosingle agent therapies, while minimizing any associated toxicity. Thesecombinations may also increase the efficacy of the conventional agentwithout increasing the associated toxicity. Preferred combinationtherapies include the administration of a compound of this inventionwith AZT, 3TC, ddI, ddC, d4T or other reverse transcriptase inhibitors.

Alternatively, the compounds of this invention may also beco-administered with other HIV protease inhibitors such as Ro 31-8959(Saquinavir; Roche), L-735,524(Indinavir; Merck), AG-1343 (Nelfinavir;Agouron), A-84538 (Ritonavir; Abbott), ABT-378/r (Lopinavir; Abbott),and VX-478 (Amprenavir; Glaxo) to increase the effect of therapy orprophylaxis against various viral mutants or members of other HIV quasispecies.

We prefer administering the compounds of this invention as single agentsor in combination with retroviral reverse transcriptase inhibitors, orother HIV aspartyl protease inhibitors. We believe that theco-administration of the compounds of this invention with retroviralreverse transcriptase inhibitors or HIV aspartyl protease inhibitors mayexert a substantial synergistic effect, thereby preventing,substantially reducing, or completely eliminating viral infectivity andits associated symptoms.

The compounds of this invention can also be administered in combinationwith immunomodulators (e.g., bropirimine, anti-human alpha interferonantibody, IL-2, GM-CSF, methionine enkephalin, interferon alpha,diethyldithiocarbamate sodium, tumor necrosis factor, naltrexone andrEPO) antibiotics (e.g., pentamidine isethionate) or vaccines to preventor combat infection and disease associated with HIV infection, such asAIDS and ARC.

When the compounds of this invention are administered in combinationtherapies with other agents, they may be administered sequentially orconcurrently to the patient. Alternatively, pharmaceutical orprophylactic compositions according to this invention may be comprisedof a combination of an aspartyl protease inhibitor of this invention andanother therapeutic or prophylactic agent.

Although this invention focuses on the use of the compounds disclosedherein for preventing and treating HIV infection, the compounds of thisinvention can also be used as inhibitory agents for other viruses thatdepend on similar aspartyl proteases for obligatory events in their lifecycle. These viruses include, but are not limited to, retrovirusescausing AIDS-like diseases such as simian immunodeficiency viruses,HIV-2, HTLV-I and HTLV-II. In addition, the compounds of this inventionmay also be used to inhibit other aspartyl proteases and, in particular,other human aspartyl proteases including renin and aspartyl proteasesthat process endothelin precursors.

Pharmaceutical compositions of this invention comprise any of thecompounds of the present invention, and pharmaceutically acceptablesalts thereof, with any pharmaceutically acceptable carrier, adjuvant orvehicle. Pharmaceutically acceptable carriers, adjuvants and vehiclesthat may be used in the pharmaceutical compositions of this inventioninclude, but are not limited to ion exchangers, alumina, aluminumstearate, lecithin, serum proteins, such as human serum albumin, buffersubstances such as phosphates, glycine, sorbic acid, potassium sorbate,partial glyceride mixtures of saturated vegetable fatty acids, water,salts or electrolytes, such as protamine sulfate, disodium hydrogenphosphate, potassium hydrogen phosphate, sodium chloride, zinc salts,colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone,cellulose-based substances, polyethyleneglycol, sodiumcarboxymethylcellulose, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat.

The pharmaceutical compositions of this invention may be administeredorally, parenterally by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir. We prefer oraladministration or administration by injection. The pharmaceuticalcompositions of this invention may contain any conventional non-toxicpharmaceutically acceptable carriers, adjuvants or vehicles. The term“parenteral” as used herein includes subcutaneous, intracutaneous,intravenous, intramuscular, intra-articular, intrasynovial,intrasternal, intrathecal, intralesional and intracranial injection orinfusion techniques.

The pharmaceutical compositions may be in the form of a sterileinjectable preparation, for example, as a sterile injectable aqueous oroleaginous suspension. This suspension may be formulated according totechniques known in the art using suitable dispersing or wetting agents(such as, for example, Tween 80) and suspending agents. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally acceptable diluent or solvent,for example, as a solution in 1,3-butanediol. Among the acceptablevehicles and solvents that may be employed are amino acid, water,Ringer's solution and isotonic sodium chloride solution. In addition,sterile, fixed oils are conventionally employed as a solvent orsuspending medium. For this purpose, any bland fixed oil may be employedincluding synthetic mono- or diglycerides. Fatty acids, such as oleicacid and its glyceride derivatives are useful in the preparation ofinjectables, as are natural pharmaceutically-acceptable oils, such asolive oil or castor oil, especially in their polyoxyethylated versions.These oil solutions or suspensions may also contain a long-chain alcoholdiluent or dispersant, such as Ph. Helv. or a similar alcohol.

The pharmaceutical compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, and aqueous suspension and solutions. Inthe case of tablets for oral use, carriers that are commonly usedinclude lactose and corn starch. Lubricating agents, such as magnesiumstearate, are also typically added. For oral administration in a capsuleform, useful diluents include lactose and dried corn starch. Whenaqueous suspensions are administered orally, the active ingredient iscombined with emulsifying and suspending agents. If desired, certainsweetening and/or flavoring and/or coloring agents may be added.

The pharmaceutical compositions of this invention may also beadministered in the form of suppositories for rectal administration.These compositions can be prepared by mixing a compound of thisinvention with a suitable non-irritating excipient which is solid atroom temperature but liquid at the rectal temperature and therefore willmelt in the rectum to release the active components. Such materialsinclude, but are not limited to, cocoa butter, beeswax, and polyethyleneglycols.

Topical administration of the pharmaceutical compositions of thisinvention is especially useful when the desired treatment involves areasor organs readily accessible by topical application. For applicationtopically to the skin, the pharmaceutical composition should beformulated with a suitable ointment containing the active componentssuspended or dissolved in a carrier. Carriers for topical administrationof the compounds of this invention include, but are not limited to,mineral oil, liquid petroleum, white petroleum, propylene glycol,polyoxyethylene or polyoxypropylene compound, emulsifying wax and water.Alternatively, the pharmaceutical compositions can be formulated with asuitable lotion or cream containing the active compound suspended ordissolved in a carrier. Suitable carriers include, but are not limitedto, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters waxcetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water. Thepharmaceutical compositions of this invention may also be topicallyapplied to the lower intestinal tract by rectal suppository formulationor in a suitable neat formulation. Topically-transdermal patches arealso included in this invention.

The pharmaceutical compositions of this invention may be administered bynasal aerosol or inhalation. Such compositions are prepared according totechniques well-known in the art of pharmaceutical formulation and maybe prepared as solutions in saline employing benzyl alcohol or othersuitable preservatives, absorption promoters to enhance bioavailability,fluorocarbons, and/or other solubilizing or dispersing agents known inthe art.

Dosage levels of between about 0.01 and about 25 mg/kg body weight perday, preferably between about 0.5 and about 25 mg/kg body weight per dayof the active ingredient compound are useful in the prevention andtreatment of viral infection, including HIV infection. Typically, thepharmaceutical compositions of this invention will be administered fromabout 1 to about 5 times per day or alternatively, as a continuousinfusion. Such administration can be used as a chronic or acute therapy.The amount of active ingredient that may be combined with the carriermaterials to produce a single dosage form will vary depending upon thepatient treated and the particular mode of administration. A typicalpreparation will contain from about 5% to about 95% active compound(w/w). Preferably, such preparations contain from about 20% to about 80%active compound.

Upon improvement of a patient's condition, a maintenance dose of acompound, composition or combination of this invention may beadministered if necessary. Subsequently, the dosage or frequency ofadministration, or both, may be reduced, as a function of the symptoms,to a level at which the improved condition is retained. When thesymptoms have been alleviated to the desired level, treatment shouldcease. Patients may, however, require intermittent treatment on along-term basis, upon any recurrence of disease symptoms.

As the skilled artisan will appreciate, lower or higher doses than thoserecited above may be required. Specific dosage and treatment regimen forany particular patient will depend upon a variety of factors, includingthe activity of the specific compound employed, the age, body weight,general health status, sex, diet, time of administration, rate ofexcretion, drug combination, the severity and course of the infection,the patient's disposition to the infection and the judgment of thetreating physician.

The compounds of this invention are also useful as commercial reagentswhich effectively bind to aspartyl proteases, particularly HIV aspartylprotease. As commercial reagents, the compounds of this invention, andtheir derivatives, may be used to block proteolysis of a target peptideby an aspartyl protease, or may be derivatized to bind to a stable resinas a tethered substrate for affinity chromatography applications. Theseand other uses which characterize commercial aspartyl proteaseinhibitors will be evident to those of ordinary skill in the art.

Enzymatic Assay for Determining the Inhibition Constant (Ki) ofSynthetic Compounds Targeting the HIV Protease

This is a fluorometric assay based on the cleavage by protease of asubstrate carrying a donor group (EDANS) and an acceptor group (DABCYL)on each side of the cleavage site, interacting together throughfluorescence resonance energy transfer (FRET) as described by Matayoshiet al. (Science 247:954-954, 1990).

After calculation of Vo and Vi, the inhibition constant (Ki) of thecompound is determined using the equation of Henderson:$\frac{Vo}{Vi} = {{1 + {\frac{\lbrack I\rbrack}{{Ki}_{app}}\quad {Where}\quad {Ki}}} = \frac{{Ki}_{app}}{\frac{1 + \lbrack S\rbrack}{Km}}}$

where

Vo=the enzyme's initial velocity

Vi=the enzyme velocity in the presence of the inhibitory compound,

[I]=inhibitor concentration,

[S]=substrate concentration,

Km=Michaelis-Menten constant and

Ki_(app)=apparent Ki

Graphs are traced and the Ki determined using GraphPad Prism software v.3.0.

The compounds listed in Tables 1 and 2 were prepared by following Scheme1, 2, 3, 4, 5, 6 or 7; the numbers of the compounds listed in the tablecorrespond to the example numbers presented in the experimental section(see examples below). The activities of the compounds are also listed inthe same tables demonstrating their potential usefulness. In Table 1 areshown compounds of formula I wherein Y, n, Cx, R₁, R₂, R₃, R₄ and R₅ areas presented in Table 1. In Table 2 are shown compounds of formula IIwherein Y, n, Cx, R₁, R₂, R₄ and R_(a) are as presented in Table 2.

In the description herein, the following abbreviations are used:

Abbreviation Meaning Ac Acetyl AcOH Acetic acid ARC AIDS-related complexAIDS Acquired Immunodeficiency Syndrome AZT 3-Azido-3-deoxythymine(Zidovudine) Bn benzyl Boc tert-Butoxycarbonyl i-Bu iso-Butyl t-Butert-Butyl CAM Cerium ammonium molybdate DABCYL4-[[4′-(dimethylamino)phenyl]azo]benzoic acid DCCDicyclohexylcarbodiimide DCE Dichloroethane DCM Dichloromethane DMAPN,N-dimethylaminopyridine DIEA N,N-Diisopropylethylamine DMFDimethylformamide DNA Deoxyribonucleic acid EDANS5-[(2′-aminoethyl)amino]naphthalene sulfonic acid EDC1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride EtOAc Ethylacetate EtOH Ethyl alcohol Fmoc 9-Fluorenylmethoxycarbonyl g Gram HIV-1,-2 Human immunodeficiency virus type 1, type 2 HOBt1-Hydroxybenzotriazole HPLC High performance liquid chromatographyHTLV-I, -II Human T-cell lymphotropic virus type I, type II IL-2Interleukin-2 Kg Kilogram LAH Lithium aluminum hydride LC-MS Liquidchromatography-mass spectrometry M Molar MeOH Methyl alcohol mgMilligram MP Melting point min Minute mol Mole mL Milliliter mmolMillimole nM Nanomolar i-Pr iso-Propyl rEPO Recombinant erythropoietinRNA Ribonucleic acid 3TC 2′,3′-Dideoxy-3-thiacytidine TFATrifluoroacetic acid H TFA Trifluoroacetic acid ammonium salt THFTetrahydrofuran Z Benzyloxycarbonyl

EXAMPLES

In order that this invention be more fully understood, the followingexamples are set forth. These examples are for the purpose ofillustration only and are not to be construed as limiting the scope ofthe invention in any way.

Materials and Methods

Analytical thin layer chromatography (TLC) was carried out with 0.25 mmsilica gel E. Merck 60 F₂₅₄ plates and eluted with the indicated solventsystems. Preparative chromatography was performed by flashchromatography, using silica gel 60 (EM Science) with the indicatedsolvent systems and positive air pressure to allow proper rate ofelution. Detection of the compounds was carried out by exposing elutedplates (analytical or preparative) to iodine, UV light and/or treatinganalytical plates with a 2% solution of p-anisaldehyde in ethanolcontaining 3% sulfuric acid and 1% acetic acid followed by heating.Alternatively, analytical plates can be treated with a 0.3% ninhydrinsolution in ethanol containing 3% acetic acid and/or a CAM solution madeof 20 g (NH₄)₆Mo₇O₂₄ and 8.3 g Ce(SO₄)₂ polyhydrate in water (750 mL)containing concentrated sulfuric acid (90 mL).

Preparative HPLC were perform on a Gilson apparatus equipped with a C18column, a215 liquid handler module and 15 mL/min capacity head pumps.The HPLC is operated with a Gilson UniPoint System Software. A solventgradient was used starting from H₂O/CH₃CN (95%:5%) to 100% CH₃CN over 25min, and 100% CH₃CN for a further 20 min to clean the column.

Unless otherwise indicated, all starting materials were purchased from acommercial source such as Aldrich Co. or Sigma Co.

Nuclear magnetic resonance (NMR) spectra were recorded on a Bruker AMX500 equipped with a reversed or QNP probe. Samples were dissolved indeuterochloroform (CDCl₃), deuteroacetone (acetone-d₆) ordeuterodimethylsulfoxide (DMSO-d₆) for data acquisition usingtetramethylsilane as internal standard. Chemical shifts (δ) areexpressed in parts per million (ppm), the coupling constants (J) areexpressed in hertz (Hz) whereas multiplicities are denoted as s forsinglet, d for doublet, dd for doublet of doublets, t for triplet, q forquartet, m for multiplet, and br s for broad singlet.

GENERAL PROCEDURES A. Preparation of N-acylated (or N-sulfonated) aminoacids (Schotten-Baumen procedure)

To a solution of an amino acid (10 mmol) in 25 mL 1N NaOH and 5 mLsaturated Na₂CO₃ (resulting solution at pH 10) was added an acylchloride (or a sulfonyl chloride, or a chloroformate) (12 mmol)dissolved in 10 mL acetone over a period of 20 min. Afterwards, thereaction mixture was stirred at room temperature for 2 h. The alkalinesolution was extracted once with ether (50 mL) and the aqueous phase wasacidified with 1N HCl to form a pasty oil. This was extracted twice with20 mL CHCl₃, and the combined organic phases were washed with 10 mL 1NHCl. The organic phase was dried over MgSO₄, filtered and evaporated toan oil which crystallized on standing. The solid was recrystallized fromeither dichloromethane, ether, hexanes or without solvent as indicatedin each specific example. The purity was evaluated by LC-MS and/or ¹HNMR and was found to be ranging from 85 to 99%.

B. Coupling reaction of N-acylated (or N-sulfonated) amino acid with theNε-NH₂ of a L-lysine derivative

Depending on the nature of the reagents, various methods were used tolink the two amino acid portions together.

a) N,N-carbonyldiimidazole Method

Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine hydrochloride (orother L-lysine derivative) (100 mg, 0.25 mmol) was weighed in theBohdahn robotic reaction vessels. The solid was then added to 1 mL 3.3MCs₂CO₃ solution to which 2 mL of THF was added. The tube was thenstirred vigorously. The resulting mixture was treated with N-acylated(or N-sulfonated) amino acid (0.3 mmol) activated byN,N-carbonyldiimidazole (0.3 mmol) dissolved in THF (1 mL). The stirringwas continued for 2 h. Afterwards, EtOAc (3 mL) was added and theorganic phase was removed. The organic phase was washed with 1N HCl andagain separated. Evacuation of the solvent gave a crude product whichwas resolved by HPLC. The yield of the reactions will be indicated ineach specific example.

b) Solid Phase Method

Preparation of Solid-phase BoundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysine

Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysine(1.51 g, 2.6 mmol) was dissolved in DCM (70 mL) containing DCC (1.5 g).The solution was stirred at room temperature for 8 h and then filtered.The filtrate was added to 5.0 g dried washed Wang resin (0.73 meq/g) towhich 150 mg N,N-dimethylaminopyridine (DMAP) was added. The suspensionwas stirred at room temperature for 12 h. Then, the resin was filteredand washed successively with DCM (100 mL, 2×), 1:1 DCM: MeOH (100 mL,3×), MeOH (50 mL, 2×) and ether (100 mL). The resin was again swollen inDCM to which acetic anhydride (20 mL) was added. It was left to standfor 3 h and then filtered and washed as above. The resulting resin wasdried at room temperature in a dessicator in vacuo. The resulting resin(5.92 g) contained 0.28 meq/g L-lysine derivative.

NB: Same preparation for solid-phase boundNα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysine(see example 28).

Deprotection

In a typical experiment, 450 mg (0.125 mmol) of resin was added to asyringe type reaction vessel with Teflon frits and stopcock. The resinwas swollen with DCM and washed after 15 min. It was treated with 30%piperidine in DMF (4 mL) and left for 15 min before being successivelywashed with DMF (5 mL, 2×), DCM (5 mL, 4×), and ether (5 mL, 4×). Thisprocess was repeated once.

Coupling

In a typical experiment, 0.5 mmol of N-acylated (or N-sulfonated) aminoacid was added to a solution of N-hydroxysuccinimide (0.5 mmol) and DCC(0.5 mmol) in DMF (3 mL). The acid was activated for 3 h and filtereddirectly into the resin containing vessel. The coupling reaction wasallowed to proceed for 12 h at room temperature. The resin was thenwashed successively with DCM, MeOH and ether as described above thendried in vacuo.

Cleavage

The dried resin was swollen with DCM, filtered and treated with 95% TFA(4 mL). The resulting mixture was stirred for a period of 3 h. Then, thesolution was filtered off and evaporated. The residue was trituratedwith ether and the pasty solid placed under high vacuum for 4 h. Thesolid was purified by preparative HPLC to give the final coupledproduct. The yield of the reactions will be indicated in each specificexample.

c) Dicyclohexylcarbodiimide (DCC) Method

Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine potassium salt (orother L-lysine potassium salt derivative) (1 mmol) was weighed in around bottom flask and was suspended in THF (20 mL). This suspension wastreated with a 1N NaOH (1.5 mL) to pH 10. In another reaction vessel, asolution of N-protected amino acid (1 mmol) in THF (25 mL), was treatedwith 115 mg of N-hydroxysuccinimide (1 mmol) and 206 mg DCC (1 mmol).The resulting mixture was stirred at room temperature for 4 h.Afterwards, the precipitate was filtered off and the filtrate was addedto the initial suspension with vigorous stirring. After 2 h, a 2 mLaliquot of water was added resulting in a clear solution. The stirringwas continued for 12 h. Then, EtOAc (50 mL) was added and the organicphase was washed successively with 1N NaOH (50 mL), with 1N HCl (50 mL)and finally with brine (50 mL). The organic phase was removed and driedwith Na₂SO₄. The solvent was evaporated and the product was purified (inwhole or in part) by preparative HPLC or by trituration with ether. Theyield of the reactions will be indicated in each specific example.

d) 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC)Method

A solution of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysinol (1.63g) in EtOAc (16 mL) was prepared (100 mg/mL, 0.29 mmol/mL). A secondsolution containing EDC (2.5 g) and HOBt (1.34 g) in DMF was prepared(0.5 mmol/mL). To Bohdan robotic test tubes were then added a series ofN-substituted amino acids (0.5 mmol) to which a 1 mL aliquot of theEDC:HOBt solution was added. After 20 min, a 1 mL aliquot of the lysinolsolution was added. The resulting solutions were stirred at roomtemperature for 6 h. Afterwards, a 5 mL aliquot of 10% citric acidaqueous solution was added to each test tube and the solutions wereextracted with EtOAc (50 mL). The organic phase were evaporated and theresidue obtained in each test tube was purified by HPLC. The yield ofthe reactions will be indicated in each specific example.

C. Removal of the N-tert-butoxycarbonyl (Boc) Group

To a series of Boc protected products (100 mg) in Bohdan test tubes wasadded 2 mL of CH₂Cl₂:TFA (1:1). Gas evolution was observed and thesolutions were stirred for 20 min. The solvents were evacuated and theresulting thick oil was triturated with cold ether. The ether wasdecanted away and the remaining products were placed in a high vacuumdesiccator for 12 h to give solid foams. The yield of the reactions willbe indicated in each specific example.

D. Sodium Salt Formation

The L-lysine product (100 mg) was dissolved in MeCN (1 mL). The solutionwas added to 5 mL H₂O to form a turbid suspension. A 1N NaOH solution (1mol eq) was slowly added to the turbid suspension which became clear.The solution was frozen solid and lyophilized to yield a white powder(100%).

E. Catalytic Hydrogenation

To a series of nitro compounds (100 mg) dissolved in argon saturatedMeOH (10 mL) was added 10% Pd/C (50 mg) followed by formic acid 98% (0.1mL). The suspensions were saturated with H₂ and kept under positivepressure using a H₂ filled balloons. After 4-6 h stirring, the H₂ waspurged out and the solutions were filtered through thin pads of celite.The clear solutions were then evacuated, triturated with ether andresolved by preparative HPLC. The yield of the reactions will beindicated in each specific example.

F. General Procedure for the Thiomidation Reaction

To a stirred solution of amide (1 mmol) in dry THF (10 mL) was addedLawesson's reagent (606 mg, 1.5 mmol). The reaction was stirredovernight then concentrated and purified by flash chromatography usinghexane/EtOAc as eluent to afford the desired thioamide.

G. General Procedure for the Reduction of esters with LiAlH₄

To a stirred solution of the ester (1 mmol) in THF was added at 0° C.LiAlH₄ (1.5 mmol). The mixture was stirred at room temperature for 3 h.The hydride excess was neutralised with HCl 1N and the reaction wasextracted with EtOAc. The organic phase was dried (MgSO₄) andconcentrated. The crude was purified by flash chromatography.

H. Substitution Reaction on an iodoacetamide Derivative

To a solution ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-iodoacetyl-L-lysine (1.0eq.) in THF (10 mL) was added DIEA (2.0-3.0 eq.) and an amine derivative(2.0-4.0 eq.). The reaction mixture was stirred overnight at roomtemperature. Then, a 2N HCl solution (2 mL) was added and the resultingmixture was extracted with EtOAc (20 mL, 3×). The organic phase wasdried MgSO₄ and evaporated to an oil. The crude material was purified bypreparative HPLC.

SPECIFIC EXAMPLES FOR THE PREPARATION OF DERIVATIVES OF GENERAL FORMULAI

The following compounds were prepared either from a L-amino acid or,when indicated, from a derivative of a D-amino acid using the proceduressummarized in schemes 1, 2, 3, 4, 5, 6 and 7.

Example 1 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-phenylalanyl]-L-lysine

Step A. Preparation of L-lysine methyl ester dihydrochloride.MeOH (J.Org. Chem. 44, 4841 (1979))

To a stirred suspension of L-lysine monohydrochloride (190.7 g, 1.08mol) in MeOH (3 L) was added (via a cannula) trimethylsilylchloride (350mL). The mixture quickly became clear and homogeneous. The solution wasstirred at reflux for 3 h and then at room temperature for 2 h. Thereaction flask was left overnight in a refrigerator cooled to −75° C.The large crystals obtained were filtered, washed with cold MeOH (100mL) and dried in vacuo for 24 h at room temperature. L-lysine methylester dihydrochloride MeOH (275.8 g) was obtained in 99.4% yield.

¹H NMR (DMSO-d₆): δ 1.36 (m, 1H), 1.45 (m, 1H), 1.58 (m, 2H), 1.81 (m,22H), 2.74 (br s, 2H), 3.11 (s, 3H), 3.72 (s, 3H), 3.94 (t, J=4.0, 1H),8.12 (br s, 3H), 8.72 (br s, 3H).

Step B. Preparation of L-α-amino-ε-caprolactam hydrochloride (J. Org.Chem. 44,4841 (1979))

Sodium methylate 58.73 g (1 mole) was dissolved in cold MeOH (1 L).About one half of this solution was cannulated into a solution ofL-lysine methyl ester dihydrochloride MeOH (132.5 g, 0.5 mole) in 1 LMeOH. The suspension was allowed to warm and dissolved. The remaindersodium methylate was added with concurrent apparition of NaCl. Themixture was then allowed to reflux for 4 h, after which 5 g of NH₄Cl wasadded. The solution then sat at RT for 18 h and was filtered throughcelite. Evaporation of the MeOH resulted in a thick opaque syrup. Theexcess NaCl was removed by redissolving the mixture in boiling glyme(100 mL, 2×), filtering through celite and evaporating in vacuo. Theresulting clear oil was taken up in ethanol and acidified with 12 N HCl.Cooling gave a mass of fine white needles which were filtered and driedin vacuo to yield 69.71 g, 85% of the title compound. MP 301-306° C.

[α]d=−24.8 (c=3.4, 1N HCl). ¹H NMR (DMSO-d₆): δ 1.17 (q, J=12.6, 1H),1.45 (q, J=12.6, 1H), 1.58 (q, J=12.6, 1H), 1.71 (d, J=12.6, 1H), 1.86(d, J=12.6, 1H), 1.94 (d, J=12.6, 1H), 3.03 (m, 1H), 3.15 (m, 1H), 4.03(d, J=12.6, 1H), 8.12 (br s, 1H), 8.22 (br s, 3H). ¹³C NMR (DMSO-d₆): δ28.2, 29.7, 29.9, 41.6, 53.4, 173.2. LC-MS: 129.1 (M+H)⁺, 99% pure.

Step C. Preparation of Nα-isobutyl-L-α-amino-ε-caprolactam

L-α-amino-ε-caprolactam (60.0 g, 0.47 mol) was dissolved indichloroethane (DCE, 100 mL) containing isobutyraldehyde (37.0 g, 0.5mole) and stirred until the heat evolved was dissipated. Then, DCE (2 L)and AcOH (35 mL) were added to the solution followed by 0.5 mole ofpowdered NaBH(OAc)₃. The slightly turbid mixture was stirred at 60° C.for 2 h, and at room temperature for 12 h. The solution was treated with1M K₂CO₃ (1 L) and stirred for a further 2 h. The DCE layer was driedwith MgSO₄, filtered and evaporated. The oil thus obtained crystallizesslowly on standing (87 g, 94.5%) and was used without furtherpurification in the next step. MP 52-54° C. A small sample was convertedto the hydrochloride salt by adding the solid to a solution of 1N HCl in95% EtOH.

¹H NMR (CDCl₃): δ 0.93 (d, J=6.5, 3H), 0.97 (d, J=6.5, 3H), 1.39 (t,J=9.8, 1H), 1.47 (m, 1H), 1.78-1.65 (m, 2H), 2.00-1.93 (m, 2H), 2.32-2.2(m, 2H), 2.38 (t, J=9.7, 1H), 3.16 (m, 3H), 6.62 (s, 1H (NH)).

Step D. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-α-amino-ε-caprolactam

The compound prepared in step C of this example (10.0 g, 51 mmol, freebase) was dissolved in DCM (100 mL) and treated withdiisopropylethylamine (10 mL) followed by freshly recrystallized4-methylbenzenesulfonyl chloride (11.4 g, 57.3 mmol). The mixture wasstirred overnight (TLC shows the reaction to be complete after 2 h). Thesolution was extracted with 1N HCl and the organic layer was dried andevaporated. Then, the residue was dissolved in boiling CHCl₃ (5 mL),diluted with hexanes (200 mL) and placed in the refrigerator for 3 h.The precipitated product was filtered off and air dried giving 15.5 g ofpure product. MP 49-51° C. ¹H NMR (CDCl₃): δ 0.74(d, J=6.2, 3H), 0.80(d, J=6.2, 3H), 1.12 (q, J=8.3, 1H), 1.56-1.73 (m, 4H), 1.84-1.87 (m,1H), 1.96-1.99 (m, 1H), 2.33 (s, 3H), 2.86-2.89 (m, 1H), 2.97-2.98 (m1H), 3.1-3.06 (m, 2H), 3.21-3.26 (m, 1H), 4.48 (d, J=10.6, 1H), 5.7 (s,1H (NH)), 7.29 (d, J=7.7, 2H), 7.59 (d, J=7.7, 2H).

Step E. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysinehydrochloride

A mixture ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-α-amino-ε-caprolactam (13.5g, 40 mmol), AcOH (4 mL) and 6N HCl (200 mL) was refluxed for 12 h untilall solids had disappeared. Afterwards, the solution was evaporated togive 11.0 g, 77% of the hydrochloride salt.

¹H NMR (DMSO-d₆): δ 0.72 (dd, J=5.8, 6.4, 6H), 1.13-1.17 (m, 2H),1.17-1.24 (m 2H), 1.42-1.48 (m, 2H), 2.3 (s, 3H), 2.67 (t, J=7.2, 2H),2.80-2.91 (m, 2H), 4.13 (t, J=7.2, 1H), 7.22 (d, J=8.5, 2H), 7.64 (d,J=8.5, 2H).

Step F. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-phenylalanyl]-L-lysine

A suspension of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysinehydrochloride (600 mg) in THF (20 mL) was treated with a 1N NaOH (1.5mL) to pH 10. A solution of commercially availableNα-(4-methylbenzenesulfonyl)-L-phenylalanine acid chloride (250 mg) indry THF (20 mL) was added to the suspension and stirred for 2 h.Afterwards, water (2 mL) was added resulting in a clear solution. Thereaction mixture was stirred for 12 h. Then, EtOAc (30 mL) was added andthe organic phase was washed with 1N HCl. The organic phase was removed.Evaporation of the solvent gave a crude product which was trituratedwith ether to yield 750 mg (76%) of the title compound.

1H NMR (CDCl₃): δ 0.77 (d, J=6.3, 3H), 0.80 (d, J=6.3, 3H), 1.00-1.11(m, 4H), 1.23-1.25 (m, 1H), 1.70-1.74 (m, 1H), 1.89-1.93 (m, 1H), 2.30(s, 3H), 2.32 (s, 3H), 2.59-2.67 (m, 2H), 2.87 and 2.93 (ABX, J=14.1,4.2, 2H), 3.85 (t, J=5.9, 1H), 3.63 (t, J=6.9, 1H), 6.90-7.10 (m, 7H),7.24 (d, J=8.0, 2H), 7.44 (d, J=8.1, 2H), 7.73 (d, J=8.1, 2H). LC-MS:656.2 (M−H)⁻, 98% pure.

Example 2 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-tryptophanyl]-L-lysine

Step A. Preparation of Nα-(4-methylbenzenesulfonyl)-L-tryptophan

L-tryptophan was reacted with 4-methylbenzenesulfonyl chloride under theconditions used in general procedure A giving the title compound whichwas recrystallised from DCM (58%).

¹H NMR (CDCl₃): δ 2.33 (s, 3H), 2.9-3.11 (m, 2H), 3.91 (t, J=7.0, 1H),6.86-7.01 (m, 3H), 7.25 (d, J=6.9, 2H), 7.34 (t, J=6.8, 1H), 7.45 (d,J=6.9, 2H), 8.15 (d, J=6.1, 1H). LC-MS: 357 (M−H)⁻, 99% pure.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-tryptophanyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (150 mg, 0.42 mmol,example 1, step E) as described in general procedure Bc usingNα-(4-methylbenzenesulfonyl)-L-tryptophan (180 mg, 0.5 mmol) which wasprepared in step A of this example. The final product was trituratedwith ether to yield 180 mg (69%) of the desired material.

¹H NMR (DMSO-d₆): δ 0.77 (d, J=6.3, 3H), 0.80 (d, J=6.3, 3H), 1.00-1.11(m, 4H), 1.32-1.35 (m, 1H), 1.70-1.74 (m, 1H), 1.83-1.88 (m, 1H), 2.26(s, 3H), 2.34 (s, 3H), 2.59-2.67 (m, 2H), 2.92 and 2.87 (ABX, J=13.1,2.8, 2H), 3.80 (t, J=6.5, 1H), 4.11 (t, J=7.2, 1H), 6.85 (t, J=7.1, 1H),7.00 (t, J=4.0, 2H), 7.10 (d, J=7.1, 2H), 7.28 (d, J=4.0, 1H), 7.33 (m,3H), 7.43 (d, J=7.1, 2H), 7.60 (d, J=7.1, 2H), 7.71 (t, J=5.4, 1H), 7.80(d, J=8.0, 1H). LC-MS: 695.2 (M−H)⁻, 99% pure.

Example 3 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-acetamidobenzenesulfonyl)-L-phenylalanyl]-L-lysine

Step A. Preparation of Nα-(4-acetamidobenzenesulfonyl)-L-phenylalanine

L-phenylalanine was reacted with 4-acetamidobenzenesulfonyl chlorideunder the conditions used in general procedure A giving the titlecompound which was recrystallised from DCM (52%).

LC-MS: 362 (M−H)⁻, 95% pure.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-acetamidobenzenesulfonyl)-L-phenylalanyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingNα-(4-acetamidobenzenesulfonyl)-L-phenylalanine (102 mg, 0.29 mmol)which was prepared in step A of this example. The final product wastriturated with ether to yield 101 mg (57%) of the desired material.

LC-MS: 699.2 (M−H)⁻, 95% pure.

Example 4 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzenesulfonyl-L-tryptophanyl)-L-lysine

Step A. Preparation of Nα-benzenesulfonyl-L-tryptophan

L-tryptophan was reacted with benzenesulfonyl chloride under theconditions used in general procedure A giving the title compound whichwas recrystallised from DCM (26%).

¹H NMR (CDCl₃): δ 2.86-3.26 (m, 2H), 3.93 (t, J=5.0, 1H), 6.92-7.00 (m,3H), 7.28 (b, J=7.0, 2H), 7.30-7.34 (m, 3H), 7.55 (d, J=6.0, 1H), 8.24(d, J=6.0, 1H). LC-MS: 343 (M−H)⁻, 99% pure.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzenesulfonyl-L-tryptophanyl)-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (1.0 g, 2.9 mmol,example 1, step E) as described in general procedure Bc usingNα-benzenesulfonyl-L-tryptophan (1.72 g, 5 mmol) which was prepared instep A of this example. The final product was triturated with ether toyield 1.7 g of the crude material. Purification of 500 mg of the crudematerial by HPLC gave 322 mg (64%) of pure adduct.

¹H NMR (CDCl₃): δ 0.78 (d, J=6.3, 3H), 0.81 (d, J=6.3, 3H), 0.94-1.03(m, 4H), 1.32-1.35 (m, 1H), 1.46-1.49 (m, 1H), 1.83-1.88 (m, 1H), 2.35(s, 3H), 2.82-2.99 (m, 4H), 3.95 (t, J=6.5, 1H), 4.21 (t, J=7.2, 1H),6.85 (t, J=4.5, 1H), 7.09 (t, J=4.5, 1H), 7.23-7.31 (m, 6H), 7.42 (t,J=4.5, 1H), 7.60 (d, J=6.8, 2H), 7.73 (d, J=6.8, 2H). LC-MS: 681.2(M−H)⁻, 99% pure.

Example 5 Preparation ofNα-isobutyl-Nα-(4-aminobenzenesulfonyl)-Nε-[N′α-(4-aminobenzenesulfonyl)-L-tryptophanyl]-L-lysine

Step A. Preparation of Nα-(4-nitrobenzenesulfonyl)-L-tryptophan

L-tryptophan was reacted with 4-nitrobenzenesulfonyl chloride under theconditions used in general procedure A givingNα-(4-nitrobenzenesulfonyl)-L-tryptophan which was recrystallised fromDCM (56%).

LC-MS: 388 (M−H)⁻, 99% pure.

Step B. Preparation ofNα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-L-α-amino-ε-caprolactam

Nα-isobutyl-L-α-amino-ε-caprolactam (example 1, step C) (4.14 g, 21.1mmol, free base) was dissolved in DCM (50 mL) and treated withdiisopropylethylamine (6.0 mL, 30 mmol) followed by freshlyrecrystallized 4-nitrobenzenesulfonyl chloride (5.09 g, 21.7 mmol). Themixture was stirred overnight (TLC shows the reaction to be completeafter 2 h). The solution was extracted with 1N HCl and the organic layerwas dried and evaporated. Then, the residue was dissolved in boilingMeOH (250 mL) and placed in the refrigerator for 3 h. The thin needlesobtained were filtered off and air dried giving 6.9 g (83%) of pureproduct. MP 152-154° C.

¹H NMR (CDCl₃): δ 0.93 (d, J=6.0, 3H), 0.96 (d, J=6.0, 3H), 1.39 (t,J=12.0, 1H), 1.65-1.85 (m, 3H), 2.08-2.18 (m, 3H), 3.06 (dd, J=14.2,8.5, 1H), 3.35 (dd, J=14.2, 8.5, 1H), 4.65 (d, J=8.7, 1H), 5.7 (s, 1H(NH)), 7.92 (d, J=8.8, 2H), 8.3 (d, J=8.8, 2H).

Step C. Preparation of Nα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-L-lysinehydrochloride

A mixture ofNα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-L-α-amino-ε-caprolactam (1.0 g,2.7 mmol), AcOH (4 mL) and 6N HCl (10 mL) was refluxed for 12 h untilall solids had disappeared. Afterwards, the solution was evaporated togive 1.12 g, 100% of the hydrochloride salt.

[α]d=−16.7 (c=0.36 in MeOH); ¹H NMR (DMSO-d₆): δ 0.79 (d, J=6.8,3H),0.86 (d, J=6.8, 3H), 1.25 (t, J=11.9, 2H), 1.32-1.28 (m 2H),1.58-1.45 (m, 2H), 1.85-1.75 (m, 2H), 2.7 (m, 3H (NH)), 2.83-2.87 (m,1H), 3.03-3.07 (m, 1H), 4.21 (t, J=10.1, 1H), 8.10 (d, J=7.9, 2H), 8.37(d, J=7.9, 2H).

Step D. Preparation ofNα-isobutyl-Nα-(4-aminobenzenesulfonyl)-Nε-[N′α-(4-aminobenzenesulfonyl)-L-tryptophanyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-L-lysine hydrochloride (200 mg,0.52 mmol, step C) as described in general procedure Bc usingNα-(4-nitrobenzenesulfonyl)-L-tryptophan (300 mg, 0.8 mmol) which wasprepared in step A of this example. The intermediate derivative wasreduced following the indications of general procedure E. The finalproduct was purified by HPLC to give 101 mg (53%) of pure adduct.

¹H NMR (DMSO-d₆): δ 0.73 (d, J=6.3, 3H), 0.75 (d, J=6.3, 3H), 1.00-1.11(m, 4H), 1.32-1.35 (m, 1H), 1.66-1.69 (m, 1H), 1.83-1.88 (m, 1H), 2.48(br s, 6H), 2.59-2.67 (m, 2H), 2.84-2.96 (m, 2H), 3.80 (t, J=6.5, 1H),4.01 (t, J=7.2, 1H), 6.46 (d, J=7.1, 2H), 6.52 (d, J=7.1, 2H), 6.85 (t,J=4.0, 1H), 7.09 (t, J=4.0, 2H), 7.28 (d, J=7.1, 1H), 7.33 (d, J=7.1,2H), 7.60 (t, J=4.0, 1H). LC-MS: 697.2 (M−H)⁻, 98% pure.

Example 6 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-nitrobenzenesulfonyl)-L-tryptophanyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingNα-(4-nitrobenzenesulfonyl)-L-tryptophan (120 mg, 0.3 mmol) which wasprepared in step A of example 5. The final product was purified by HPLCto give 66 mg (36%) of pure adduct.

LC-MS: 726.2 (M−H)⁻, 99% pure.

Example 7 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-D-phenylalanyl]-L-lysine

Step A. Preparation of Nα-(4-methylbenzenesulfonyl)-D-phenylalanine

D-phenylalanine was reacted with 4-methylbenzenesulfonyl chloride underthe conditions used in general procedure A giving the title compoundwhich was recrystallised from ether (18%).

LC-MS: 318 (M−H)⁻, 98% pure.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-D-phenylalanyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingNα-(4-methylbenzenesulfonyl)-D-phenylalanine (160 mg, 0.5 mmol) whichwas prepared in step A of this example. The final product was trituratedwith ether to yield 49 mg (29%) of the desired material.

LC-MS: 656.2 (M−H)⁻, 99% pure.

Example 8 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzenesulfonyl-L-phenylalanyl)-L-lysine

Step A. Preparation of Nα-benzenesulfonyl-L-phenylalanine

L-phenylalanine was reacted with benzenesulfonyl chloride under theconditions used in general procedure A giving the title compound whichwas recrystallised from ether (50%).

LC-MS: 349 (M−H)⁻, 95% pure.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzenesulfonyl-L-phenylalanyl)-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine hydrochloride (120 mg,0.34 mmol, example 1, step E) as described in general procedure Bc usingNα-benzenesulfonyl-L-phenylalanine (300 mg, 1.0 mmol) prepared in step Aof this example. The final product was purified by preparative HPLC toyield 107 mg (56%) of the desired material.

¹H NMR (CDCl₃): δ 0.74 (d, J=6.3, 3H), 0.80 (d, J=6.3, 3H), 1.10-1.21(m, 2H), 1.26-1.33 (m, 2H), 1.70-1.74 (m, 1H), 1.89-1.93 (m, 2H), 2.39(s, 3H), 2.79-2.89 (m, 2H), 3.85 (t, J=5.9, 1H), 4.29 (t, J=6.9, 1H),6.90 (d, J=6.2, 2H), 7.08-7.29 (m, 6H), 7.35 (t, J=6.2, 2H), 7.44 (d,J=8.1, 2H), 7.73 (d, J=8.1, 2H). LC-MS: 642.2 (M−H)⁻, 99% pure.

Example 9 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-chlorobenzenesulfonyl)-L-phenylalanyl]-L-lysine

Step A. Preparation of Nα-(4-chlorobenzenesulfonyl)-L-phenylalanine

L-phenylalanine was reacted with 4-chlorobenzenesulfonyl chloride underthe conditions used in general procedure A giving the title compoundwhich was recrystallised neat (30%).

LC-MS: 338 (M−H)⁻, 98% pure.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-chlorobenzenesulfonyl)-L-phenylalanyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingNα-(4-chlorobenzenesulfonyl)-L-phenylalanine (89 mg, 0.3 mmol) preparedin step A of this example. The final product was purified by preparativeHPLC to yield 56 mg (33%) of the desired material.

¹H NMR (DMSO-d₆): δ 0.74 (d, J=6.3, 3H), 0.80 (d, J=6.3, 3H), 1.00-1.11(m, 4H), 1.23-1.25 (m, 1H), 1.70-1.74 (m, 1H), 1.89-1.93 (m, 1H), 2.32(s, 3H), 2.59-2.67 (m, 2H), 2.84 and 2.95 (ABX, J=14.1, 6.8, 2H), 3.85(t, J=5.9, 1H), 4.11 (t, J=6.9, 1H), 7.02-7.21 (m, 7H), 7.24 (d, J=8.0,1H), 7.54 (d, J=8.1, 2H), 7.73 (d, J=8.1, 2H), 8.07 (d, J=6.4, 1H).LC-MS: 677.2 (M−H)⁻, 99% pure.

Example 10 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-nitrobenzenesulfonyl)-L-phenylalanyl]-L-lysine

Step A. Preparation of Nα-(4-nitrobenzenesulfonyl)-L-phenylalanine

L-phenylalanine was reacted with 4-nitrobenzenesulfonyl chloride underthe conditions used in general procedure A giving the title compoundwhich was recrystallised in ether (37%).

LC-MS: 349 (M−H)⁻, 98% pure.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-nitrobenzenesulfonyl)-L-phenylalanyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingNα-(4-nitrobenzenesulfonyl)-L-phenylalanine (125 mg, 0.5 mmol) preparedin step A of this example. The final product was purified by preparativeHPLC to yield 91 mg (52%) of the desired material.

LC-MS: 687.2 (M−H)⁻, 98% pure.

Example 11 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-tyrosyl]-L-lysine

Step A. Preparation of Nα-(4-methylbenzenesulfonyl)-L-tyrosine

L-tyrosine was reacted with 4-methylbenzenesulfonyl chloride under theconditions used in general procedure A giving the title compound whichwas recrystallised neat (15%).

LC-MS: 334 (M−H)⁻, 95% pure.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-tyrosyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingNα-(4-methylbenzenesulfonyl)-L-tyrosine (160 mg, 0.5 mmol) prepared instep A of this example. The final product was purified by preparativeHPLC to yield 78 mg (46%) of the desired material.

LC-MS: 672.2 (M−H)⁻, 99% pure.

Example 12 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-aminobenzenesulfonyl)-L-tryptophanyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine hydrochloride (200 mg,0.59 mmol, example 1, step E) as described in general procedure Bc usingNα-(4-nitrobenzenesulfonyl)-L-tryptophan (300 mg, 0.75 mmol) prepared instep A of example 5. The intermediate,Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-nitrobenzenesulfonyl)-L-tryptophanyl]-L-lysine,was reduced following the conditions of general procedure E. The finalproduct was purified by preparative HPLC to yield 266 mg (58%) of thedesired material.

¹H NMR (DMSO-d₆): δ 0.74 (d, J=6.3, 3H), 0.80 (d, J=6.3, 3H), 1.00-1.11(m, 4H), 1.32-1.35 (m, 1H), 1.70-1.74 (m, 1H), 1.83-1.88 (m, 1H), 2.34(s, 3H), 2.59-2.67 (m, 2H), 2.84-2.96 (m, 2H), 3.86 (t, J=6.5, 1H), 4.13(t, J=7.2, 1H), 6.85 (t, J=4.0, 1H), 7.00 (t, J=4.0, 2H), 7.10 (d,J=7.1, 2H), 7.28 (d, J=4.0, 1H), 7.33 (m, 3H), 7.43 (d, J=7.1, 2H), 7.60(d, J=7.0, 2H), 7.71 (t, J=4.1, 1H), 7.80 (d, J=8.0, 1H). LC-MS: 725.2(M−H)⁻, 98% pure.

Example 13 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-nitrobenzenesulfonyl)-L-alanyl]-L-lysine

Step A. Preparation of Nα-(4-nitrobenzenesulfonyl)-L-alanine

L-alanine was reacted with 4-nitrobenzenesulfonyl chloride under theconditions used in general procedure A giving the title compound whichwas recrystallised neat (9%). This compound was used without furtherpurification in the next step.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-nitrobenzenesulfonyl)-L-alanyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine hydrochloride (100 mg,0.29 mmol, example 1, step E) as described in general procedure Bc usingNα-(4-nitrobenzenesulfonyl)-L-alanine (140 mg, 0.5 mmol) prepared instep A of this example. The final product was purified by preparativeHPLC to yield 10 mg (6.5%) of the desired material.

LC-MS: 611.2 (M−H)⁻, 99% pure.

Example 14 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(benzenesulfonyl)-L-norvalyl]-L-lysine

Step A. Preparation of Nα-benzenesulfonyl-L-norvaline

L-norvaline was reacted with benzenesulfonyl chloride under theconditions used in general procedure A giving the title compound whichwas recrystallised neat (33%). This compound was used without furtherpurification in the next step.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(benzenesulfonyl)-L-norvalyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step B) as described in general procedure Bc usingNα-benzenesulfonyl-L-norvaline (120 mg, 0.5 mmol) prepared in step A ofthis example. The final product was purified by preparative HPLC toyield 15 mg (10%) of the desired material.

LC-MS: 594.3 (M−H)⁻, 99% pure.

Example 15 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(benzenesulfonyl)-L-norleucyl]-L-lysine

Step A. Preparation of Nα-benzenesulfonyl-L-norleucine

L-norleucine was reacted with benzenesulfonyl chloride under theconditions used in general procedure A giving the title compound whichwas recrystallised neat (25%). This compound was used without furtherpurification in the next step.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(benzenesulfonyl)-L-norleucyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingNα-benzenesulfonyl-L-norleucine (125 mg, 0.5 mmol) prepared in step A ofthis example. The final product was purified by preparative HPLC toyield 13 mg (8.5%) of the desired material.

LC-MS: 608.3 (M−H)⁻, 99% pure.

Example 16 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-nitrobenzenesulfonyl)-L-leucyl]-L-lysine

Step A. Preparation of Nα-(4-nitrobenzenesulfonyl)-L-leucine

L-leucine was reacted with 4-nitrobenzenesulfonyl chloride under theconditions used in general procedure A giving the title compound whichwas recrystallised neat (66%). This compound was used without furtherpurification in the next step.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-nitrobenzenesulfonyl)-L-leucyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingNα-(4-nitrobenzenesulfonyl)-L-leucine (150 mg, 0.5 mmol) prepared instep A of this example. The final product was purified by preparativeHPLC to yield 28 mg (17%) of the desired material.

LC-MS: 653.3 (M−H)⁻, 99% pure.

Example 17 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(benzenesulfonyl)-4-trans-hydroxy-L-prolyl]-L-lysine

Step A. Preparation of Nα-benzenesulfonyl-4-trans-hydroxy-L-proline

4-trans-hydroxy-L-proline was reacted with benzenesulfonyl chlorideunder the conditions used in general procedure A giving the titlecompound which was recrystallised neat (21%). This compound was usedwithout further purification in the next step.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(benzenesulfonyl)-4-trans-hydroxy-L-prolyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingNα-benzenesulfonyl-4-trans-hydroxy-L-proline (130 mg, 0.5 mmol) preparedin step A of this example. The final product was purified by preparativeHPLC to yield 9 mg (6%) of the desired material.

LC-MS: 608.3 (M−H)⁻, 99% pure.

Example 18 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-fluorobenzenesulfonyl)-L-phenylalanyl]-L-lysine

Step A. Preparation of Nα-(4-fluorobenzenesulfonyl)-L-phenylalanine

L-phenylalanine was reacted with 4-fluorobenzenesulfonyl chloride underthe conditions used in general procedure A giving the title compoundwhich was recrystallised in ether (40%).

LC-MS: 322 (M−H)⁻, 99% pure.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-fluorobenzenesulfonyl)-L-phenylalanyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingNα-(4-fluorobenzenesulfonyl)-L-phenylalanine (160 mg,0.5 mmol) preparedin step A of this example. The final product was purified by preparativeBPLC to yield 87 mg (52%) of the desired material.

¹H NMR (DMSO-d₆): δ 0.74 (d, J=6.3, 3H), 0.80 (d, J=6.3, 3H), 1.00-1.11(m, 4H), 1.23-1.25 (m, 1H), 1.70-1.74 (m, 1H), 1.89-1.93 (m, 1H), 2.32(s, 3H), 2.65 (m, 2H), 2.85 and 2.95 (ABX, J=16.1 7.1, 2H), 3.88 (t,J=6.0, 1H), 4.11 (t, J=6.9, 1H), 7.02-7.21 (m, 7H), 7.24 (d, J=8.0, 1H),7.54 (d, J=8.1, 2H), 7.73 (d, J=8.1, 2H), 8.15 (d, J=6.6, 1H). LC-MS:660.2 (M−H)⁻, 99% pure.

Example 19 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(2-naphthylsulfonyl)-L-phenylalanyl]-L-lysine

Step A. Preparation of Nα-(2-naphthylsulfonyl)-L-phenylalanine

L-phenylalanine was reacted with 2-naphthylsulfonyl chloride under theconditions used in general procedure A giving the title compound whichwas recrystallised in ether (51%).

LC-MS: 352 (M−H)⁻, 95% pure.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(2-naphthylsulfonyl)-L-phenylalanyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingNα-(2-naphthylsulfonyl)-L-phenylalanine (175 mg, 0.5 mmol) prepared instep A of this example. The final product was purified by preparativeHPLC to yield 55 mg (32%) of the desired material.

LC-MS: 692.3 (M−H)⁻, 99% pure.

Example 20 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-bromobenzenesulfonyl)-L-phenylalanyl]-L-lysine

Step A. Preparation of Nα-(4-bromobenzenesulfonyl)-L-phenylalanine

L-phenylalanine was reacted with 4-bromobenzenesulfonyl chloride underthe conditions used in general procedure A giving the title compoundwhich was recrystallised in ether (18%).

LC-MS: 383 (M−H)⁻, 99% pure.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-bromobenzenesulfonyl)-L-phenylalanyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingNα-(4-bromobenzenesulfonyl)-L-phenylalanine (190 mg, 0.5 mmol) preparedin step A of this example. The final product was purified by preparativeHPLC to yield 56 mg (30%) of the desired material.

¹H NMR (DMSO-d₆): δ 0.74 (d, J=6.3, 3H), 0.80 (d, J=6.3, 3H), 1.00-1.11(m, 4H), 1.23-1.25 (m, 1H), 1.70-1.74 (m, 1H), 1.89-1.93 (m, 1H), 2.32(s, 3H), 2.60-2.68 (m, 2H), 2.86 and 2.97(ABX, J=14.1, 7.1, 2H), 3.96(t, J=6.1, 1H),4.11 (t, J=6.9, 1H), 6.99-7.22 (m, 7H), 7.24 (d, J=8.0,1H), 7.54 (d, J=8.1, 2H), 7.73 (d, J=8.1, 2H), 8.11 (d, J=6.5, 1H).LC-MS: 721.2 (M−H)⁻, 99% pure.

Example 21 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-glycyl]-L-lysine

Step A. Preparation of Nα-(4-methylbenzenesulfonyl)-glycine

Glycine was reacted with 4-methylbenzenesulfonyl chloride under theconditions used in general procedure A giving the title compound whichwas recrystallised neat (60%).

LC-MS: 228 (M−H)⁻, 99% pure.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine by following theindications of general example Bc using commercially availableN-(9-fluorenylmethoxycarbonyloxy)succinimide instead of the usualcombination of reactants i.e. N-protected amino acid,N-hydroxysuccinimide and DCC.

¹H NMR (CDCl₃): 0.79 (d, J=7.1, 3H), 0.81 (d, J=7.1, 3H), 1.12-1.25 (m,2H), 1.30-1.40 (m, 2H), 1.42-1.50 (m, 2H), 1.78-1.90 (m, 2H), 2.36 (s,3H), 2.85 (m, 2H), 2.88 and 3.04 (ABX, J=14.3, 7.3, 2H), 4.16-4.21 (m,2H), 4.28 (d, J=7.0, 2H), 7.30-7.42 (m, 6H), 7.60 (m, 4H), 7.88 (d,J=7.5, 2H), 12.69 (br s, 1H).

Step C. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-glycyl]-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysine(step B) as described in general procedure Bb usingNα-(4-methylbenzenesulfonyl)-glycine (110 mg,0.5 mmol) prepared in stepA of this example. The final product was purified by preparative HPLC toyield 30 mg (42%) of the desired material.

¹H NMR (CDCl₃): δ 0.73 (d, J=6.9, 6H), 1.23-1.25 (m, 2H), 1.45-1.52 (m,3H), 1.89-1.99 (m, 2H), 2.32 (s, 6H), 2.94-3.03 (m, 2H), 3.16 (m, 2H),3.55 (t, J=5.9, 1H), 4.27 (t, J=7.2, 1H), 7.26 (d, J=8.1, 4H), 7.73 (d,J=8.1, 4H). LC-MS: 566.5 (M−H)⁻, 85% pure.

Example 22 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzenesulfonyl-L-leucyl]-L-lysine

Step A. Preparation of Nα-benzenesulfonyl-L-leucine

L-leucine was reacted with benzenesulfonyl chloride under the conditionsused in general procedure A giving the title compound which wasrecrystallised neat (31%). This compound was used without furtherpurification in the next step.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzenesulfonyl-L-leucyl]-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysineas described in general procedure Bb using Nα-benzenesulfonyl-L-leucine(130 mg, 0.5 mmol) prepared in step A of this example. The final productwas purified by preparative HPLC to yield 30 mg (39%) of the desiredmaterial.

¹H NMR (CDCl₃): δ 0.72 (d, J=6.0, 3H), 0.75 (d, J=6.0, 3H), 0.78-0.81(m, 6H), 1.20-1.22 (m, 2H), 1.32-1.34 (m, 2H), 1.52-1.55 (m, 1H),1.78-2.04 (m, 3H), 2.32 (s, 3H), 2.81-3.01 (m, 4H), 3.56 (t, J=5.2, 1H),4.25 (t, J=6.0, 1H), 7.21-7.29 (m, 2H), 7.42-7.45 (m, 2H), 7.52 (t,J=6.1, 1H), 7.71 (d, J=7.8, 2H), 7.81 (d, J=7.8, 2H). LC-MS: 608.2(M−H)⁻, 90% pure.

Example 23 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-trifluoromethylbenzenesulfonyl)-L-phenylalanyl]-L-lysine

Step A. Preparation ofNα-(4-trifluoromethylbenzenesulfonyl)-L-phenylalanine

L-phenylalanine was reacted with 4-trifluoromethylbenzenesulfonylchloride under the conditions used in general procedure A giving thetitle compound which was recrystallised neat (18%).

LC-MS: 369 (M−H)⁻, 98% pure.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-trifluoromethylbenzenesulfonyl)-L-phenylalanyl]-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysineas described in general procedure Bb usingNα-(4-trifluoromethylbenzenesulfonyl)-L-phenylalanine (180 mg, 0.5 mmol)prepared in step A of this example. The final product was purified bypreparative HPLC to yield 50 mg (56%) of the desired material.

LC-MS: 710.2 (M−H)⁻, 80% pure.

Example 24 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(2-thiophenesulfonyl)-L-phenylalanyl]-L-lysine

Step A. Preparation of Nα-(2-thiophenesulfonyl)-L-phenylalanine

L-phenylalanine was reacted with 2-thiophenesulfonyl chloride under theconditions used in general procedure A giving the title compound whichwas recrystallised from ether (93%).

LC-MS: 310 (M−H)⁻, 99% pure.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(2-thiophenesulfonyl)-L-phenylalanyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine hydrochloride (250 mg,0.61 mmol, example 1, step E) as described in general procedure Bc usingNα-(2-thiophenesulfonyl)-L-phenylalanine (155 mg, 0.5 mmol) which wasprepared in step A of this example. The crude material was purified byHPLC to give 272 mg (66%) of pure adduct.

¹H NMR (DMSO-d₆): δ 0.77 (d, J=6.3, 3H), 0.80 (d, J=6.3, 3H), 1.00-1.11(m, 4H), 1.32-1.35 (m, 1H), 1.70-1.74 (m, 1H), 1.83-1.88 (m, 1H), 2.34(s, 3H), 2.59-2.67 (m, 2H), 2.92 and 2.96 (ABX, J=16.5, 7.1, 2H), 3.90(t, J=6.5, 1H), 4.11 (t, J=7.2, 1H), 7.00 (t, J=4.0, 1H), 7.10-7.21 (m,5H), 7.30 (d, J=4.0, 1H), 7.34 (d, J=8.1, 2H), 7.63 (d, J=8.1, 2H), 7.77(d, J=4.0, 1H), 7.81 (t, J=5.3, 1H), 8.22 (d, J=8.1, 1H). LC-MS: 648.5(M−H)⁻, 99% pure.

Example 25 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzenesulfonyl)-L-asparagyl]-L-lysine

Step A. Preparation of Nα-benzenesulfonyl-L-asparagine

L-asparagine was reacted with benzenesulfonyl chloride under theconditions used in general procedure A giving the title compound whichwas recrystallised neat (29%). This compound was used without furtherpurification in the next step.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzenesulfonyl)-L-asparagyl]-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysineas described in general procedure Bb usingNα-benzenesulfonyl-L-asparagine (350mg, 1.0 mmol) prepared in step A ofthis example. The final product was purified by preparative HPLC toyield 70 mg (91%) of the desired material.

¹H NMR (CDCl₃): δ 0.78 (d, J=6.3, 3H), 0.81 (d, J=6.3, 3H), 0.94-1.13(m, 4H), 1.32-1.35 (m, 1H), 1.66-1.69 (m, 1H), 1.83-1.88 (m, 1H),2.10-2.29 (m, 2H), 2.35 (s, 3H), 2.38 (s, 3H), 2.82 and 2.99 (ABX,J=12.6, 8.1, 2H), 4.00 (t, J=6.5, 1H), 4.11 (t, J=7.2, 1H), 7.21 (d,J=7.9, 2H), 7.32 (d, J=7.9, 2H), 7.55-7.64 (m, 3H), 7.77 (d, J=7.8, 2H).LC-MS: 609.1 (M−H)⁻, 99% pure.

Example 26 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-4-nitrophenylalanyl]-L-lysine

Step A. Preparation ofNα-(4-methylbenzenesulfonyl)-L-4-nitrophenylalanine

L-4-nitrophenylalanine was reacted with 4-methylbenzenesulfonyl chlorideunder the conditions used in general procedure A giving the titlecompound which was recrystallised neat (14%).

LC-MS: 364 (M−H)⁻, 98% pure.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-4-nitrophenylalanyl]-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysineas described in general procedure Bb usingNα-(4-methylbenzenesulfonyl)-L-4-nitrophenylalanine (180 mg, 0.5 mmol)prepared in step A of this example. The final product was purified bypreparative HPLC to yield 25 mg (28%) of the desired material.

LC-MS: 701.1 (M−H)⁻, 95% pure.

Example 27 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-phenylglycyl]-L-lysine

Step A. Preparation of Nα-(4-methylbenzenesulfonyl)-L-phenylglycine

L-phenylglycine was reacted with 4-methylbenzenesulfonyl chloride underthe conditions used in general procedure A giving the title compoundwhich was recrystallised neat (21%).

LC-MS: 288 (M−H)⁻, 99% pure.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-phenylglycyl]-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysineas described in general procedure Bb usingNα-(4-methylbenzenesulfonyl)-L-phenylglycine (150 mg, 0.5 mmol) preparedin step A of this example. The final product was purified by preparativeHPLC to yield 55 mg (68%) of the desired material.

LC-MS: 642.1 (M−H)⁻, 95% pure.

Example 28 Preparation ofNα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-Nε-[N′α-(4-acetamidobenzenesulfonyl)-L-phenylalanyl]-L-lysine

Step A. Preparation ofNα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysine

Nα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-L-lysine hydrochloride (1 mmol,example 5, step C) was partially dissolved in K₂CO₃ (1M)/THF/CH₃CN (4mL/4 mL/4 mL). To this suspension was addedN-(9-fluorenylmethoxycarbonyloxy) succinimide (371 mg, 1.10 mmol). Thereaction turned slowly to colorless and was left stirring for 1 h. HCl(1M) was added until acidic pH and the reaction mixture was extractedtwice with EtOAc. The combined organic layers were washed with brine,dried over MgSO₄ and concentrated. The residue was purified by flashchromatography eluting with a mixture of hexane/EtOAc containing 0.4%AcOH to yield 88% of the title compound which was used without furtherpurification in the next step.

Step B. Preparation ofNα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-Nε-[N′α-(4-acetamidobenzenesulfonyl)-L-phenylalanyl]-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysine(step A) as described in general procedure Bb usingNα-(4-acetamidobenzenesulfonyl)-L-phenylalanine (400 mg, 1.2 mmol) whichwas prepared in step A of example 3. The final product was purified bypreparative HPLC to yield 55 mg (60%) of the desired material.

LC-MS: 730.1 (M−H)⁻, 95% pure.

Example 29 Preparation ofNα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-Nε-[N′α-(2-thiophenesulfonyl)-L-phenylalanyl]-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysineas described in general procedure Bb usingNα-(2-thiophenesulfonyl)-L-phenylalanine (300 mg, 1.2 mmol) which wasprepared in step A of example 24. The final product was purified bypreparative HPLC to yield 46 mg (54%) of the desired material.

LC-MS: 679.0 (M−H)⁻, 95% pure.

Example 30 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-acetyl-L-phenylalanyl)-L-lysine

Step A. Preparation of Nα-acetyl-L-phenylalanine

L-phenylalanine was reacted with acetyl chloride under the conditionsused in general procedure A giving the title compound which wasrecrystallised from ether (97%). This compound is also commerciallyavailable.

LC-MS: 206 (M−H)⁻, 99% pure.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-acetyl-L-phenylalanyl)-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (150 mg, 0.42 mmol,example 1, step E) as described in general procedure Bc usingNα-acetyl-L-phenylalanine (207 mg, 1.0 mmol) which was prepared in stepA of this example. The final product was purified by preparative HPLC toyield 121 mg (59%) of the desired material.

¹H NMR (CDCl₃): δ 0.83 (d, J=6.9, 6H), 1.08-1.11 (m, 2H), 1.23-1.25 (m,2H), 1.45-1.52 (m, 1H), 1.89-1.99 (m and s (1.90), 5H), 2.32 (s, 3H),2.94-3.09 (m, 6H), 4.23 (t, J=5.9), 4.61 (m, 1H), 7.09-7.26 (m, 7H),7.73 (d, J=8.1, 2H). LC-MS: 544.2 (M−H)⁻, 99% pure.

Example 31 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzyloxycarbonyl-L-phenylalanyl)-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysineas described in general procedure Bb using commercially availableNα-benzyloxycarbonyl-L-phenylalanine (300 mg, 1.2 mmol). The finalproduct was purified by preparative HPLC to yield 33 mg (37%) of thedesired material.

LC-MS: 636.2 (M−H)⁻, 99% pure.

Example 32 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-seryl]-L-lysine

Step A. Preparation of Nα-(4-methylbenzenesulfonyl)-L-serine

L-serine was reacted with 4-methylbenzenesulfonyl chloride under theconditions used in general procedure A giving the title compound whichwas recrystallised neat (44%). This compound was used without furtherpurification in the next step.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-seryl]-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysineas described in general procedure Bb usingNα-(4-methylbenzenesulfonyl)-L-serine (150 mg, 1.2 mmol) prepared instep A of this example. The final product was purified by preparativeHPLC to yield 35 mg (46%) of the desired material.

¹H NMR (CDCl₃): δ 0.78-0.82 (m, 6H), 1.18-1.22 (m, 2H), 1.38-1.41 (m,2H), 1.50-1.52 (m, 1H), 1.79-1.96 (m, 2H), 2.32 (s, 6H), 2.85-2.97 (m,2H), 3.06-3.19 (m, 2H), 3.82 (br s, 1H), 4.23 (t, J=6.9, 1H), 7.25 (d,J=8.0, 4H), 7.70 (d, J=8.0, 4H). LC-MS: 596.1 (M−H)⁻, 95% pure.

Example 33 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-cyclohexylalanyl]-L-lysine

Step A. Preparation of Nα-(4-methylbenzenesulfonyl)-L-cyclohexylalanine

L-cyclohexylalanine was reacted with 4-methylbenzenesulfonyl chlorideunder the conditions used in general procedure A giving the titlecompound which was recrystallised neat (14%).

LC-MS: 324 (M−H)⁻, 95% pure.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-cyclohexylalanyl]-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysineas described in general procedure Bb usingNα-(4-methylbenzenesulfonyl)-L-cyclohexylalanine (350 mg, 1.2 mmol)prepared in step A of this example. The final product was purified bypreparative HPLC to yield 19 mg (22%) of the desired material.

¹H NMR (CDCl₃): δ 0.73-1.73 (m, 24H), 1.89-1.99 (m, 2H), 2.39 (s, 6H),2.94-3.19 (m, 4H), 3.65 (t, J=5.9, 1H), 4.33 (t, J=7.2, 1H), 7.26 (d,J=8.1, 4H), 7.73 (d, J=8.1, 4H), LC-MS: 662.2 (M−H)⁻, 95% pure.

Example 34 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-glutaminyl]-L-lysine

Step A. Preparation of Nα-(4-methylbenzenesulfonyl)-L-glutamine

L-glutamine was reacted with 4-methylbenzenesulfonyl chloride under theconditions used in general procedure A giving the title compound whichwas recrystallised neat (11%). This compound was used without furtherpurification in the next step.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-glutaminyl]-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysineas described in general procedure Bb usingNα-(4-methylbenzenesulfonyl)-L-glutamine (360 mg, 1.2 mmol) prepared instep A of this example. The final product was purified by preparativeHPLC to yield 18 mg (21%) of the desired material.

LC-MS: 637.2 (M−H)⁻, 80% pure.

Example 35 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-2-thiophenylalanyl]-L-lysine

Step A. Preparation ofNα-(4-methylbenzenesulfonyl)-L-2-thiophenylalanine

L-2-thiophenylalanine was reacted with 4-methylbenzenesulfonyl chlorideunder the conditions used in general procedure A giving the titlecompound which was recrystallised neat (33%). This compound was usedwithout further purification in the next step.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-2-thiophenylalanyl]-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysineas described in general procedure Bb usingNα-(4-methylbenzenesulfonyl)-L-2-thiophenylalanine (390 mg, 1.2 mmol)prepared in step A of this example. The final product was purified bypreparative HPLC to yield 15 mg (18%) of the desired material.

LC-MS: 662.1 (M−H)⁻, 85% pure.

Example 36 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(9-fluorenylmethoxycarbonyl)-L-seryl]-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysineas described in general procedure Bb using commercially availableNα-(9-fluorenylmethoxycarbonyl)-L-serine (390 mg, 1.2 mmol). The finalproduct was purified by preparative HPLC to yield 15 mg (18%) of thedesired material.

LC-MS: 664.3 (M−H)⁻, 95% pure.

Example 37 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(9-fluorenylmethoxycarbonyl)-L-cyclohexylalanyl]-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysineas described in general procedure Bb using commercially availableNα-(9-fluorenylmethoxycarbonyl)-L-cyclohexylalanine (470 mg, 1.2 mmol).The final product was purified by preparative HPLC to yield 11 mg (12%)of the desired material.

LC-MS: 730.6 (M−H)⁻, 95% pure.

Example 38 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-glutamyl]-L-lysine

Step A. Preparation of Nα-(4-methylbenzenesulfonyl)-L-glutamic acid

L-glutamic acid was reacted with 4-methylbenzenesulfonyl chloride underthe conditions used in general procedure A giving the title derivative(20%) which was used without purification in the next step.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-glutamyl]-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysineas described in general procedure Bb usingNα-(4-methylbenzenesulfonyl)-L-glutamic acid (360 mg, 1.2 mmol) preparedin step A of this example. The final product was purified by preparativeHPLC to yield 20 mg (25%) of the desired material.

LC-MS: 640.3 (M+H)⁺, 99% pure.

Example 39 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-lysyl]-L-lysine

Step A. Preparation ofNα-(4-methylbenzenesulfonyl)-Nε-tert-butoxycarbonyl-L-lysine

Nε-tert-butoxycarbonyl-L-lysine was reacted with 4-methylbenzenesulfonylchloride under the conditions used in general procedure A giving thetitle derivative which was used without purification in the next step.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-lysyl]-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysineas described in general procedure Bb usingNα-(4-methylbenzenesulfonyl)-Nε-tert-butoxycarbonyl-L-lysine (360 mg,1.2 mmol) prepared in step A of this example. The final product waspurified by preparative HPLC to yield 10 mg (12%) of the desiredmaterial.

LC-MS: 637.2 (M−H)⁻, 99% pure.

Example 40 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-O-benzyl-L-seryl]-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysine(200 mg, 0.1 mmol) in a similar fashion to general procedure Bb usingcommercially available Nα-(9-fluorenylmethoxycarbonyl)-O-benzyl-L-serine(100 mg, 0.24 mmol) with DCC (100 mg, 0.48 mmol) and HOBt (50 mg, 0.37mmol) as the activating reagents. The intermediate adduct was againdeprotected and coupled with 4-methylbenzenesulfonyl chloride in CH₂Cl₂before being cleaved from the resin with TFA. The final product waspurified by preparative HPLC to yield 30 mg (42%) of the desiredmaterial.

¹H NMR (DMSO-d₆): δ 0.78 (d, J=6.5, 3H), 0.82 (d, J=6.6, 3H), 1.15-1.45(m, 5H), 1.84 (m, 1H), 1.95 (m, 1H), 2.34 (s, 3H), 2.36 (s, 3H), 2.79(m, 2H), 2.91 (m, 2H), 3.42 (t, J=5.7, 2H), 3.92 (t, J=6.1, 1H), 4.04(m, 1H), 4.37 (d, J=5.7, 2H), 7.21 (d, J=7.3, 2H), 7.23-7.33 (m, 7H),7.65 (d, J=8.2, 2H), 7.71 (d, J=7.7, 2H), 7.93 (t, J≅4.0, 1H), 7.95 (brs, 1H). LC-MS: 6.88 (M+H)⁺, 99% pure.

Example 41 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-aspartyl]-L-lysine

Step A. Preparation of Nα-(4-methylbenzenesulfonyl)-L-aspartic acid

L-aspartic acid was reacted with 4-methylbenzenesulfonyl chloride underthe conditions used in general procedure A giving the title derivative(40%) which was used without purification in the next step.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-aspartyl]-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysineas described in general procedure Bb usingNα-(4-methylbenzenesulfonyl)-L-aspartic acid (340 mg, 1.2 mmol) preparedin step A of this example. The final product was purified by preparativeHPLC to yield 20 mg (26%) of the desired material.

LC-MS: 624.1 (M−H)⁻, 99% pure.

Example 42 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-3-(2-thianaphthyl)-alanyl]-L-lysine

Step A. Preparation of4-methylbenzenesulfonyl-L-3-(2-thianaphthyl)-alanine

L-3-(2-thianaphthyl)-alanine was reacted with 4-methylbenzenesulfonylchloride under the conditions used in general procedure A giving thetitle derivative which was recrystallised neat (34%). This compound wasused without further purification in the next step.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-3-(2-thianaphthyl)-alanyl]-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysineas described in general procedure Bb using4-methylbenzenesulfonyl-L-3-(2-thianaphthyl)-alanine (450 mg, 1.2 mmol)prepared in step A of this example. The final product was purified bypreparative HPLC to yield 25 mg (28%) of the desired material.

¹H NMR (CDCl₃): δ 0.85-0.89 (m, 6H), 1.08-1.15 (m, 2H), 1.32-1.36 (m,2H), 1.50-1.53 (m, 2H), 1.85-1.89 (m, 2H), 2.24 (s, 3H), 2.96-3.12 (m,4H), 3.84-3.86 (m, 1H), 4.22 (t, J=5.4, 1H), 6.90 (d, J=6.8, 2H), 7.06(s, 1H), 7.19-7.28 (m, 8H), 7.46 (d, J=6.8, 1H), 7.74 (d, J=6.8, 2H).LC-MS: 714.2 (M+H)⁺, 99% pure.

Example 43 Preparation ofNα-(4-aminobenzenesulfonyl)-Nα-isobutyl-Nε-[N′α-(4-methylbenzenesulfonyl)-L-tryptophanyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-L-lysine hydrochloride (350 mg,1.0 mmol, example 5, step C) as described in general procedure Bc usingNα-(4-methylbenzenesulfonyl)-L-tryptophan (335 mg, 1.1 mmol) which wasprepared in step A of example 2. The final product was purified bypreparative HPLC to yield 650 mg (90%) ofNα-(4-nitrobenzenesulfonyl)-Nα-isobutyl-Nε-[N′α-(4-methylbenzenesulfonyl)-L-tryptophanyl]-L-lysineThe latter derivative (300 mg) was hydrogenolysed following theindications of general procedure E. Purification by HPLC gave thedesired material (235 mg, 75%).

¹H NMR (DMSO-d₆): δ 0.74 (d, J=6.3, 3H), 0.80 (d, J=6.3, 3H), 1.00-1.11(m, 4H), 1.32-1.35 (m, 1H), 1.70-1.74 (m, 1H), 1.83-1.88 (m, 1H), 2.34(s, 3H), 2.59-2.67 (m, 2H), 2.84-2.96 (m, 2H), 3.80 (t, J=6.5, 1H), 4.11(t, J=7.2, 1H), 6.85 (t, J=5.2, 1H), 7.00 (t, J=4.0, 2H), 7.10 (d,J=5.3, 2H), 7.28 (d, J=4.0, 1H), 7.33 (m, 3H), 7.43 (d, J=6.9, 2H), 7.60(d, J=6.9, 2H), 7.71 (t, J=6.9, 1H), 7.80 (d, J=8.0, 1H). LC-MS: 696.8(M−H)⁻, 99% pure.

Example 44 Preparation ofNα-(4-aminobenzenesulfonyl)-Nα-isobutyl-Nε-(Nα-benzenesulfonyl-L-phenylalanyl)-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-L-lysine (385 mg, 1.0 mmol,example 5, step C) as described in general procedure Bc usingNα-benzenesulfonyl-L-phenylalanine (335 mg, 1.1 mmol) which was preparedin step A of example 8. The final product was purified by preparativeHPLC to yield 550 mg (85%) ofNα-(4-nitrobenzenesulfonyl)-Nα-isobutyl-Nε-(Nα-benzenesulfonyl)-L-phenylalanyl)-L-lysine.The latter derivative (200 mg) was hydrogenolysed following theindications of general procedure E. Purification by HPLC gave thedesired material (129 mg, 65%).

¹H NMR (CDCl₃): δ 0.75 (d, J=7.1, 3H), 0.88 (d, J=7.1, 3H), 1.0-1.1 (m,2H), 1.16-1.23 (m, 1H), 1.56-1.58 (m, 1H), 1.72-1.73 (m, 1H), 1.82-1.91(m, 1H), 2.79-2.96 (m, 2H), 3.86 7 (t, J=6.2, 1H), 4.09 (t, J=6.1, 1H),5.59 (s, 1H), 6.55 (d, J=7.5, 2H), 7.05 (d, J=7.1, 2H), 7.11-7.19 (m,4H), 7.38 (d, J=6.9, 2H), 7.42 (t, J=7.1, 2H), 7.51 (d, J=7.1, 2H), 8.01(d, J=7.1, 1H). LC-MS: 643.2 (M−H)⁻, 99% pure.

Example 45 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-tryptophanyl]-L-lysine2,3-dihydroxypropylester

A solution ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[Nα-(4-methylbenzenesulfonyl)-L-tryptophanyl]-L-lysine(product of example 2, 70 mg, 0.1 mmol) in DMF (1 mL) was treated withglycerol (100 mg) and EDC (100 mg, 0.5 mmol) and stirred overnight. Thesolution is then poured in 5% citric acid and extracted with EtOAc (5mL). The solvent was evaporated and the residue was purified bypreparative HPLC to yield 30 mg (40%) of the desired ester.

LC-MS: 769.3 (M−H)⁻, 99% pure.

Example 46 Preparation ofNα-(4-aminobenzenesulfonyl)-Nα-isobutyl-Nε-[N′α-(2-thiophenesulfonyl)-L-phenylalanyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-L-lysine hydrochloride (400 mg,1.03 mmol, example 5, step C) as described in general procedure Bc usingNα-(2-thiophenesulfonyl)-L-phenylalanine (311 mg, 1.2 mmol) which wasprepared in step A of example 24. The final product was purified bypreparative HPLC to yield 550 mg (80%) ofNα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-Nε-[N′α-(2-thiophenesulfonyl)-L-phenylalanyl]-L-lysine.The latter derivative was hydrogenolysed following the indications ofgeneral procedure E. Purification by HPLC gave the desired material (400mg, 65%).

¹H NMR (DMSO-d₆): δ 0.78 (q, J=6.9, 6H), 1.00-1.11 (m, 4H), 1.32-1.35(m, 1H), 1.70-1.74 (m, 1H), 1.83-1.88 (m, 1H), 2.59-2.67 (m, 2H),2.74-2.96 (m, 4H), 3.90 (t, J=6.5, 1H), 4.11 (t, J=7.2, 1H), 6.50 (d,J=8.1, 2H), 6.95 (t, J=4.0, 1H), 7.10-7.21 (m, 5H), 7.30 (d, J=4.0, 1H),7.34 (d, J=8.1, 2H), 7.77 (d, J=4.0, 1H), 7.81 (t, J=5.3, 1H), 8.22 (d,J=8.1, 1H). LC-MS: 649.8 (M−H)⁻, 98% pure.

Example 47 Preparation ofNα-(4-aminobenzenesulfonyl)-Nα-isobutyl-Nε-[N′α-(4-methylbenzenesulfonyl)-L-asparagyl]-L-lysine

Step A. Preparation Nα-(4-methylbenzenesulfonyl)-L-asparagine

L-asparagine was reacted with 4-methylbenzenesulfonyl chloride under theconditions used in general procedure A giving the title compound whichwas recrystallised from ether (40%).

¹H NMR (CDCl₃): δ 2.20-2.23 (m, 1H), 2.33 (s, 3H), 2.42-2.50 (m, 1H),4.00-4.03 (br s, 1H), 6.8 (s, 1H), 7.26 (s, 2H), 7.51 (s, 2H).

Step B. Preparation ofNα-(4-aminobenzenesulfonyl)-Nα-isobutyl-Nε-[N′α-(4-methylbenzenesulfonyl)-L-asparagyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-L-lysine hydrochloride (400 mg,1.03 mmol, example 5, step C) as described in general procedure Bc usingNα-(4-methylbenzenesulfonyl)-L-asparagine (286 mg, 1.0 mmol) which wasprepared in step A this example. The final product,Nα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-asparagyl]-L-lysinewas subsequently hydrogenolysed following the indications of generalprocedure E. Purification by HPLC gave the desired material (185 mg,76%).

¹H NMR (CDCl₃): δ 0.78 (d, J=6.3, 3H), 0.81 (d, J=6.3, 3H), 0.94-1.13(m, 4H), 1.32-1.35 (m, 1H), 1.66-1.69 (m, 1H), 1.83-1.88 (m, 1H),2.10-2.29 (m, 2H), 2.31 (s, 3H), 2.82-2.99 (m, 2H), 3.95 (t, J=6.5, 1H),4.11 (t, J=7.2, 1H), 6.80 (d, J=8.0, 2H), 7.19 (d, J=7.9, 2H), 7.32 (d,J=7.9, 2H), 7.64 (d, J=8.0, 2H). LC-MS: 624.8 (M−H)⁻, 98% pure.

Example 48 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzyloxycarbonyl-L-asparagyl)-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysineas described in general procedure Bb using commercially availableNα-benzyloxycarbonyl-L-asparagine (319 mg, 1.2 mmol). The final productwas purified by preparative HPLC to yield 46 mg (41%) of the desiredmaterial.

LC-MS: 603.1 (M−H)⁻, 98% pure.

Example 49 Preparation ofNα-(4-aminobenzenesulfonyl)-Nα-isobutyl-Nε-[N′α-(2-thiophenesulfonyl)-L-phenylalanyl]-L-lysinehydrazide

A solution ofNα-(4-aminobenzenesulfonyl)-Nα-isobutyl-Nε-[N′α-(2-thiophenesulfonyl)-L-phenylalanyl]-L-lysine(650 mg, 1.0 mmol, example 46) dissolved in EtOAc (20 mL) was treatedwith p-nitrophenol (139 mg, 1.0 mmol) and DCC (206 mg, 1.0 mmol) and wasleft in a refrigerator overnight. Afterwards, the precipitate wasfiltered off through celite and the solvent evaporated. The cruderesidue was used without further purification. A portion of the residue(55 mg, 0.072 mmol) was added to a solution of hydrazine hydrate inethanol (1M, 10 mL). The resulting solution was stirred for 3 h beforeevaporation of the solvent. The residue was purified by preparative HPLCto yield 25 mg, 30% of the desired material.

LC-MS: 663.1 (M−H)⁻, 95% pure.

Example 50 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzoyl-L-phenylalanyl)-L-lysine

Step A. Preparation of Nα-benzoyl-L-phenylalanine

L-phenylalanine was reacted with benzoyl chloride under the conditionsused in general procedure A giving the title compound which wasrecrystallised from ether (60%).

¹H NMR (CDCl₃): δ 3.21-3.39 (m, 2H), 5.09 (q, J=6.6, 1H), 7.10-7.27 (m,5H), 7.42 (t, J=6.9, 2H), 7.5 (t, J=7.0, 1H), 7.68 (d, J=6.9, 2H).LC-MS: 268 (M−H)⁻, 99% pure.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzoyl-L-phenylalanyl)-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysineas described in general procedure Bb using Nα-benzoyl-L-phenylalanine(321 mg, 1.2 mmol). The final product was purified by preparative HPLCto yield 44 mg (58%) of the desired material.

¹H NMR (CDCl₃): δ 0.83 (d, J=6.9, 6H), 1.08-1.11 (m, 2H), 1.23-1.25 (m,2H), 1.45-1.52 (m, 1H), 1.89-1.99 (m, 5H), 2.31 (s, 3H), 2.94-3.19 (m,6H), 4.24 (t, J=6.9, 1H), 4.89 (t, J=5.9, 1H), 7.19-7.26 (m, 7H), 7.30(t, J=7.9, 2H), 7.46 (t, J=7.8, 1H), 7.73-7.82 (m, 4H). LC-MS: 606.2(M−H)⁻, 98% pure.

Example 51 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-morpholinecarbonyl)-L-phenylalanyl]-L-lysine

Step A. Preparation of Nα-(4-morpholinecarbonyl)-L-phenylalanine

L-phenylalanine was reacted with 4-morpholinecarbonyl chloride under theconditions used in general procedure A giving the title compound whichwas recrystallised from ether (69%).

¹H NMR (CDCl₃): δ 3.11-3.39 (m, 6H), 3.57 (s, 4H), 4,65 (q, J=6.6, 1H),7.10-7.27 (m, 5H).

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-morpholinecarbonyl)-L-phenylalanyl]-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysineas described in general procedure Bb usingNα-(4-morpholinecarbonyl)-L-phenylalanine (330 mg, 1.2 mmol). The finalproduct was purified by preparative HPLC to yield 41 mg (53%) of thedesired material.

LC-MS: 615.2 (M−H)⁻, 95% pure.

Example 52 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-pivaloyl-L-phenylalanyl)-L-lysine

Step A. Preparation of Nα-pivaloyl-L-phenylalanine

L-phenylalanine was reacted with pivaloyl chloride under the conditionsused in general procedure A giving the title compound which wasrecrystallised from hexanes (40%).

¹H NMR (CDCl₃): δ 1.16 (s 9H), 3.12-3.31 (m, 2H), 4.85 (t, J=5.1, 1H),7.11-7.32 (m, 5H).

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-pivaloyl-L-phenylalanyl)-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysineas described in general procedure Bb using Nα-pivaloyl-L-phenylalanine(300 mg, 1.2 mmol). The final product was purified by preparative HPLCto yield 49 mg (66%) of the desired material.

LC-MS: 586.1 (M−H)⁻, 95% pure.

Example 53 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(2-thiophenesulfonyl)-L-tryptophanyl]-L-lysine

Step A. Preparation of Nα-(2-thiophenesulfonyl)-L-tryptophan

L-tryptophan was reacted with 2-thiophenesulfonyl chloride under theconditions used in general procedure A giving the title compound whichwas recrystallised from DCM (32%).

¹H NMR (CDCl₃): δ 2.9-3.11 (m, 2H), 3.91 (t, J=7.0, 1H), 6.86-7.01 (m,4H), 7.21-7.3 (m, 3H), 7.70 (s, 1H), 8.38 (s, 1H).

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(2-thiophenesulfonyl)-L-tryptophanyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingNα-(2-thiophenesulfonyl)-L-tryptophan (90 mg, 0.3 mmol) which wasprepared in step A of this example. The final product was trituratedwith ether to yield 12 mg (6%) of the desired material.

LC-MS: 687.2 (M−H)⁻, 85% pure.

Example 54 Preparation ofNα-isobutyl-Nα-(2-thiophenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-phenylalanyl]-L-lysine

Step A. Preparation ofNα-isobutyl-Nα-(2-thiophenesulfonyl)-L-α-amino-ε-caprolactam

Nα-isobutyl-L-α-amino-ε-caprolactam (example 1, step C) (2.56 g, 14.0mmol, free base) was dissolved in DCM (50 mL) and treated withdiisopropylethylamine (4.0 mL, 20 mmol) followed by freshlyrecrystallized 2-thiophenenesulfonyl chloride (2.56 g, 14.0 mmol). Themixture was stirred overnight (TLC shows the reaction to be completeafter 2 h). The solution was extracted with 1N HCl and the organic layerwas dried and evaporated. Then, the residue was dissolved in boilingMeOH (150 mL) and placed in the refrigerator for 3 h. The thin needlesobtained were filtered off and air dried giving 3.6 g (78%) of pureproduct. This compound was used without further purification in the nextstep.

Step B. Preparation of Nα-isobutyl-Nα-(2-thiophenesulfonyl)-L-lysinehydrochloride

A mixture ofNα-isobutyl-Nα-(2-thiophenesulfonyl)-L-α-amino-ε-caprolactam (3.5 g,10.1 mmol), AcOH (12 mL) and 6N HCl (50 mL) was refluxed for 6 h untilall solids had disappeared. Afterwards, the solution was evaporated togive 2.8 g, 71% of the hydrochloride salt.

¹H NMR (DMSO-d₆): δ 0.72 (dd, J=5.8, 6.4, 6H), 1.13-1.17 (m, 2H),1.42-1.46 (m 2H), 1.79-1.87 (m, 2H), 2.67 (t, J=7.2, 2H), 2.80-2.91 (m,2H), 4.13 (t, J=7.2, 1H), 7.11 (t, J=5.1, 1H), 7.50 (d, J=5.5, 1H), 7.85(d, J=5.6, 1H).

Step C. Preparation ofNα-isobutyl-Nα-(2-thiophenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-phenylalanyl]-L-lysine

A suspension of Nα-isobutyl-Nα-(2-thiophenesulfonyl)-L-lysinehydrochloride (150 mg, 0.5 mmol), in THF (20 mL) was treated with a 1NNaOH (0.5 mL) to pH 10. A solution of commercially availableNα-(4-methylbenzenesulfonyl)-L-phenylalanine acid chloride (150 mg, 0.4mmol), in dry THF (10 mL) was added to the suspension and stirred for 4h. Afterwards, water (2 mL) was added resulting in a clear solution.Then, EtOAc (30 mL) was added and the organic phase was washed with 1NHCl. The organic phase was removed. Evaporation of the solvent gave acrude product which was purified by preparative HPLC to yield 150 mg(57%) of the title compound.

¹H NMR (DMSO-d₆): δ 0.79 (d, J=6.3, 3H), 0.82 (d, J=6.3, 3H), 1.00-1.11(m, 4H), 1.23-1.25 (m, 1H), 1.80-1.84 (m, 1H), 1.89-1.98 (m, 1H), 2.32(s, 3H), 2.59-2.67 (m, 2H), 2.84 and 2.95 (ABX, J=15.5, 4.8, 2H), 3.88(t, J=6.0, 1H), 4.21 (t, J=6.9, 1H), 7.04 (d, J=7.6, 2H), 7.12-7.21 (m,5H), 7.35-7.44 (m, 3H), 7.87 (d, J=7.61, 2H). LC-MS: 648.5 (M−H)⁻, 98%pure.

Example 55 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-isobutyl-N′α-(4-methylbenzenesulfonyl)-glycyl]-L-lysine

Step A. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-glycine

The title compound was prepared in a three-step sequence from tert-butylbromoacetate. Initially, tert-butyl bromoacetate (1.91 g, 10 mmol)dissolved in isobutylamine (60 mmol) was stirred at room temperature for2 h. The reaction mixture was filtered and the excess isobutylamine wasdistilled yielding 80% of pure N-isobutyl glycine tert-butyl ester.Secondly, the intermediate was reacted with 4-methylbenzenesulfonylchloride (8 mmol) as described for the preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-α-amino-ε-caprolactampresented in example 1 (step D). In this case, triethylamine was usedinstead of diisopropylethylamine. Thirdly, deprotection of thetert-butyl ester with TFA provided the final product quantitatively (twolast steps).

¹H NMR (CDCl₃): δ 0.80 (br s, 6H), 1.75-1.82 (m, 1H), 2.32 (s, 3H),2.85-2.9 (br s, 2H), 3.83 (s, 2H), 7.26 (d, J=7.9, 2H), 7.6 (d, J=7.9,2H). LC-MS: 286 (M−H)⁻, 99% pure.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-isobutyl-N′α-(4-methylbenzenesulfonyl)-glycyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-glycine (90 mg, 0.3 mmol) whichwas prepared in step A of this example. The final product was trituratedwith ether to yield 40 mg (21%) of the desired material.

LC-MS: 623.8 (M−H)⁻, 95% pure.

Example 56 Preparation ofNα-(4-aminobenzenesulfonyl)-Nα-isobutyl-Nε-[N′α-(4-acetylaminobenzenesulfonyl)-L-phenylalanyl]-L-lysine

The title compound was prepared fromNα-(4-aminobenzenesulfonyl)-Nα-isobutyl-L-lysine (100 mg, 0.29 mmol,example 166, step B) as described in general procedure Bc usingNα-(4-acetamidobenzenesulfonyl)-L-phenylalanine (110 mg, 0.3 mmol) whichwas prepared in step A of example 3. The final product was purified byHPLC to give 50 mg (23%) of pure adduct.

LC-MS: 700.8 (M−H)⁻, 95% pure.

Example 57 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(2-thiophenesulfonyl)-L-phenylalanyl]-L-lysinemethyl ester

The title compound was prepared by treatingNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(2-thiophenesulfonyl)-L-phenylalanyl]-L-lysine(60 mg, 0.1 mmol, example 24) dissolved in MeOH (2 mL) with DCC (1 eq.).The reaction mixture was stirred at room temperature for a period of 2h. Filtration and evaporation of the solvent followed by HPLCpurification gave the desired methyl ester (15 mg, 22%).

LC-MS: 662.1 (M−H)⁻, 99% pure.

Example 58 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(2-thiophenesulfonyl)-L-phenylalanyl]-L-lysinamide

A solution ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(2-thiophenesulfonyl)-L-phenylalanyl]-L-lysine(650 mg, 1.0 mmol, example 24) dissolved in EtOAc (10 mL) was treatedwith p-nitrophenol (139 mg, 1.0 mmol) and DCC (206 mg, 1.0 mmol) and wasleft in a refrigerator overnight. Afterwards, the precipitate wasfiltered off through celite and the solvent evaporated. The crudeintermediate was used without further purification in the next step. Aportion of the intermediate (25 mg, 0.03 mmol) was added to a solutionof ammonia in ethanol (1M, 10 mL). The resulting solution was stirredfor 3 h before evaporation of the solvent. The residue was purified bypreparative HPLC to yield 2.6 mg, 11% of the desired material.

LC-MS: 647.2 (M−H)⁻, 99% pure.

Example 59 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(2-thiophenesulfonyl)-L-phenylalanyl]-L-lysineN-hydroxylamide

A portion of the crude intermediate of example 58 above, (25 mg, 0.03mmol) was added to a solution of hydroxylamine in ethanol (1M, 10 mL).The resulting solution was stirred for 3 h before evaporation of thesolvent. The residue was purified by preparative HPLC to yield 4.0 mg,18% of the desired material.

LC-MS: 663.1 (M−H)⁻, 99% pure.

Example 60 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(2-thiophenesulfonyl)-L-phenylalanyl]-L-lysineethanolamide

A portion of the crude intermediate of example 58 above, (25 mg, 0.03mmol) was added to a solution of ethanolamine in ethanol (1M, 10 mL).The resulting solution was stirred for 3 h before evaporation of thesolvent. The residue was purified by preparative HPLC to yield 15 mg,21% of the desired material.

LC-MS: 691.2 (M−H)⁻, 99% pure.

Example 61 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-asparagyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingNα-(4-methylbenzenesulfonyl)-L-asparagine (90 mg, 0.3 mmol) which wasprepared in step A of example 47. The final product was triturated withether to yield 95 mg (15%) of the desired material.

1H NMR (CDCl₃): δ 0.78 (d, J=6.3, 3H), 0.81 (d, J=6.3, 3H), 0.99-1.03(m, 4H), 1.32-1.35 (m, 1H), 1.66-1.69 (m, 1H), 1.83-1.88 (m, 1H),2.10-2.29 (m, 2H), 2.35 (s, 3H), 2.38 (s, 3H), 2.82 and 2.99 (m, 2H),3.95 (t, J=6.5, 1H), 4.11 (t, J=7.2, 1H), 7.19 (d, J=7.9, 2H), 7.32 (d,J=7.9, 2H), 7.55-7.64 (m, 4H). LC-MS: 625.8 (M+H)⁺, 97% pure.

Example 62 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-pivaloyl-L-asparagyl)-L-lysine

Step A. Preparation of Nα-pivaloyl-L-asparagine

L-asparagine was reacted with pivaloyl chloride under the conditionsused in general procedure A giving the title compound which wasrecrystallised neat (80%). This compound was used without furtherpurification in the next step.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-pivaloyl-L-asparagyl)-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingNα-pivaloyl-L-asparagine (65 mg, 0.3 mmol) which was prepared in step Athis example. The final product was purified by preparative HPLC toyield 21 mg (12%) of the desired material.

LC-MS: 555.7 (M+H)⁺, 90% pure.

Example 63 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzoyl-L-asparagyl)-L-lysine

Step A. Preparation of Nα-benzoyl-L-asparagine

L-asparagine was reacted with benzoyl chloride under the conditions usedin general procedure A giving the title compound which wasrecrystallised neat (90%). This compound was used without furtherpurification in the next step.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzoyl-L-asparagyl)-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingNα-benzoyl-L-asparagine (70 mg, 0.3 mmol) which was prepared in step Athis example. The final product was purified by preparative HPLC toyield 14 mg (9%) of the desired material.

LC-MS: 575.2 (M+H)⁺, 99% pure.

Example 64 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(2-thiophenesulfonyl)-L-phenylalanyl]-L-lysinehydrazide

A portion of the crude intermediate of example 58 above, (50 mg, 0.06mmol) was added to a solution of hydrazine in ethanol (1M, 10 mL). Theresulting solution was stirred for 3 h before evaporation of thesolvent. The residue was purified by preparative HPLC to yield 25 mg,60% of the desired material.

LC-MS: 664.2 (M−H)⁻, 99% pure.

Example 65 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[1,2,3,4-tetrahydroisoquinoline-N′-(4-methylbenzenesulfonyl)-3-carbonyl]-L-lysine

This particular preparation is based in scheme 4 of this invention.

Step A. Preparation of Nα-isobutyl-Nε-benzyloxycarbonyl-L-lysine methylester

To a stirred solution of commercially availableNε-benzyloxycarbonyl-L-lysine methyl ester hydrochloride (9.92 g, 30mmol), AcOH (6 mL) and NaCNBH₃ (33 mmol) in MeOH (250 mL) at 0° C. wasadded a solution of isobutyraldehyde (3.01 mL, 33 mmol) in MeOH (80 mL).The solution was warmed to room temperature and stirred for 2 h. Asaturated solution of K₂CO₃ (150 mL) was added and the solution wasdecanted from the solid and coevaporated on vacuo. The residue waspartitioned between EtOAc (300 mL) and H₂O (200 mL). The organic layerwas washed with K₂CO₃ (1M) and with brine, then dried and concentrated.The crude was used in the next step without further purification.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-benzyloxycarbonyl-L-lysinemethyl ester

To a stirred solution of Nα-isobutyl-Nε-benzyloxycarbonyl-L-lysinemethyl ester (336 mg, 1 mmol) in CH₂Cl₂ (2 mL) was added4-methylbenzenesulfonyl chloride (286 mg, 1.5 mmol) and triethylamine(174 μL, 1 mmol). The reaction mixture was allowed to stir for 3 days,then it was diluted with 1N HCl and extracted with CH₂Cl₂. The organiclayer was dried (MgSO₄) and concentrated. The crude was flashchromatographed using hexane/EtOAc as eluent to obtain the correspondingsulfonamide.

Yield: 71% (steps A and B)

¹H NMR (DMSO-d₆): δ 0.84 (d, J=7.2, 3H), 0.86 (d, J=6.3, 3H), 1.30-1.68(m, 5H), 1.88-2.00 (m, 2H), 2.42 (s, 3H), 2.92 and 3.00 (ABX, J=14.7,8.2, 2H), 3.18 (m, 2H), 3.50 (s, 3H), 4.40 (t, J=7.4, 1H), 4.78 (br s,1H), 5.11 (s, 2H), 7.27-7.71 (m, 9H).

Step C. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysinemethyl ester

Pd/C 10% (120 mg) was added to a solution of the above sulfonamide (491mg, 1 mmol) in EtOAc/MeOH (3 mL/3 mL). The suspension was flushed withH₂ and maintained under H₂ pressure until complete consumption of thestarting material. The insoluble material was filtered off, and thefiltrate was concentrated under reduced pressure to give the desiredamine in quantitative yield. This compound was used without purificationin the next step.

Step D. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[1,2,3,4-tetrahydroisoquinoline-N′-(tert-butoxycarbonyl)-3-carbonyl]-L-lysinemethyl ester

To a stirred solution of the above crude amine in THF/K₂CO₃(1M) (3 mL/3mL) was added1,2,3,4-tetrahydroisoquinoline-N′-(tert-butoxycarbonyl)-3-carboxylicacid N-hydroxysuccinimide ester (451 mg, 1.2 mmol). The reaction mixturewas stirred overnight then diluted with 1N HCl and extracted with EtOAc.The organic layer was dried (MgSO₄) and concentrated. The crude waspurified by flash chromatography using hexane/EtOAc as the eluent toafford the desired product.

Step E. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[1,2,3,4-tetrahydroisoquinoline-N′-(tert-butoxycarbonyl)-3-carbonyl]-L-lysine

The above ester (314 mg, 0.5 mmol) was dissolved in THF/MeOH (2 mL/1mL), to which was added NaOH (0.6 mmol). The reaction mixture wasstirred until complete consumption of the starting ester, then dilutedwith 1N HCl until acidic pH and extracted with EtOAc. The organic phasewas dried (MgSO₄) and concentrated to give the desired acid inquantitative yield.

Overall yield: 62% (steps C, D and E)

¹H NMR (DMSO-d₆): δ 0.80 (d, J=6.5, 3H), 0.83 (d, J=6.6, 3H), 1.00-1.30(m, 4H), 1.35 (s, 9H), 1.42 (m, 1H), 1.70-1.92 (m, 2H), 2.35 (s, 3H),2.80-3.10 (m, 6H), 4.12 (m, 1H), 4.35 (m, 1H), 4.50-4.65 (m, 2H), 7.15(m, 4H), 7.35 (d, J=8.0, 2H), 7.65 (d, J=8.0, 2H), 7.78 (m, 1H).

Step F. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[1,2,3,4-tetrahydroisoquinoline-3-carbonyl]-L-lysinetrifluoroacetic acid salt

The title product was prepared by treating a solution ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[1,2,3,4-tetrahydroisoquinoline-N′-(tert-butoxycarbonyl)-3-carbonyl]-L-lysine(614 mg, 1 mmol, step E) in CH₂Cl₂ (5 mL) with TFA (3 mL) for 3 h. Theammonium salt was isolated in quantitative yield.

¹H NMR (DMSO-d₆): δ 0.80 (d, J=6.5, 3H), 0.83 (d, J=6.8, 3H), 1.22-1.57(m, 5H), 1.82-1.98 (m, 2H), 2.38 (s, 3H), 2.88-3.22 (m, 5H), 3.30 (d,J=16.5 1H), 4.10-4.40 (m, 4H), 7.25 (m, 4H), 7.40 (d, J=7.4, 2H), 7.69(d, J=7.5, 2H), 8.60 (s, 1H), 9.40 (br s, 1H), 9.63 (br s, 1H).

Step G. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[1,2,3,4-tetrahydroisoquinoline-N′-(4-methylbenzenesulfonyl)-3-carbonyl]-L-lysine

The final product was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[1,2,3,4-tetrahydroisoquinoline-3-carbonyl]-L-lysinetrifluoroacetic acid salt (step F above) following the indications ofstep B of this example. The desired material was obtained in 71% yield.

¹H NMR (DMSO-d₆): δ 0.80 (d, J=6.5, 3H), 0.83 (d, J=6.8, 3H), 1.00-1.40(m, 5H), 1.68-1.90 (m, 2H), 2.33 (s, 3H), 2.35 (s, 3H), 2.70-3.00 (m,6H), 4.12 (m, 1H), 4.42-4.53 (m, 3H), 7.10-7.40 (m, 8H), 7.63 (d, J=8.2,4H), 7.90 (m, 1H), 12.70 (br s, 1H).

Example 66 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-D-phenylalanyl]-D-lysine

The title compound was prepared in the same manner as in the previousexample (example 65) using Nα-isobutyl-Nε-benzyloxycarbonyl-D-lysinemethyl ester and 4-methylbenzenesulfonyl-D-phenylalanine as the startingmaterials.

¹H NMR (DMSO-d₆): δ 0.80 (d, J=6.5, 3H), 0.83 (d, J=6.8, 3H), 1.00-1.12(m, 4H), 1.35-1.45 (m, 1H), 1.70-1.80 (m, 1H), 1.85-1.95 (m, 1H), 2.32(s, 3H), 2.60-2.80 (m, 4H), 2.85 and 2.97 (ABX, J=14.5, 7.5, 2H), 3.90(m, 1H), 4.15 (t, J=5.0, 1H), 7.10 (d, J=7.3, 2H), 7.12-7.25 (m, 5H),7.36 (d, J=7.5, 2H), 7.50 (d, J=8.0, 2H), 7.68 (d, J=7.5, 2H), 7.75 (t,J=5.0, 1H), 7.92 (d, J=9.2, 1H).

Example 67 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-phenylalanyl]-D-lysine

The title compound was prepared in the same manner as in the example 65using Nα-isobutyl-Nε-benzyloxycarbonyl-D-lysine methyl ester and4-methylbenzenesulfonyl-L-phenylalanine as the starting materials.

¹H NMR (CDCl₃): δ 0.77 (d, J=6.3, 3H), 0.80 (d, J=6.3, 3H), 1.00-1.11(m, 4H), 1.23-1.25 (m, 1H), 1.70-1.74 (m, 1H), 1.89-1.93 (m, 1H), 2.30(s, 3H), 2.32 (s, 3H), 2.59-2.67 (m, 2H), 2.87 and 2.93 (ABX, J=14.1,4.2, 2H), 3.85 (t, J=5.9, 1H), 3.63 (t, J=6.9, 1H), 6.90-7.10 (m, 7H),7.24 (d, J=8.0, 2H), 7.44 (d, J=8.1, 2H), 7.73 (d, J=8.1, 2H). clExample 68

Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-phenylalanyl]-L-lysinemethyl ester

To a stirred solution ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine methyl ester (369 mg,1 mmol, example 65, step C) in THF/K₂CO₃(1M) (3 mL/3 mL) was addedNα-(4-methylbenzenesulfonyl)-L-phenylalanine N-hydroxysuccinimide ester(500 mg, 1.2 mmol). The reaction mixture was stirred overnight, thendiluted with 1N HCl and extracted with EtOAc. The organic layer wasdried (MgSO₄) and concentrated. The crude was purified by flashchromatography using hexane/EtOAc as eluent to afford the desiredproduct (77% yield).

¹H NMR (DMSO-d₆): δ0.83 (d, J=7.0, 3H), 0.86 (d, J=6.8, 3H), 1.22-1.50(m, 4H), 1.60 (m, 1H), 1.80-1.95 (m, 2H), 2.40 (s, 3H), 2.42 (s, 3H),2.85-3.05 (m, 4H), 3.12 (m, 2H), 3.50 (s, 3H), 3.88 (m, 3H), 3.49 (t,J=5.0, 1H), 5.22 (m, 1H), 6.42 (t, J=5.0, 1H), 6.96 (d, J=8.0, 2H),7.12-7.20 (m, 5H), 7.30 (d, J=8.0, 2H), 7.51 (d, J=7.5, 2H), 7.72 (d,J=7.8, 2H).

Example 69 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-tryptophanyl]-L-lysinemethyl ester

To a stirred solution ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine methyl ester (369 mg,1 mmol, example 65, step C) in THF/K₂CO₃ (1M) (3 mL/3 mL) was addedNα-(4-methylbenzenesulfonyl)-L-tryptophan N-hydroxysuccinimide ester(547 mg, 1.2 mmol). The reaction mixture was stirred overnight, thendiluted with 1N HCl and extracted with EtOAc. The organic layer wasdried (MgSO₄) and concentrated. The crude was purified by flashchromatography using hexane/EtOAc as eluent to afford the desiredproduct (71% yield).

¹H NMR (DMSO-d₆): δ 0.77 (t, J=7.5, 6H), 1.00-1.10 (m, 4H), 1.40 (m,1H), 1.70 (m, 1H), 1.85 (m, 1H), 2.29 (s, 3H), 2.38 (s, 3H), 2.87-3.00(m, 3H), 3.44 (s, 3H), 3.85 (m, 1H), 4.24 (t, J=7.3, 1H), 5.74 (s, 2H),6.88-7.10 (m, 3H), 7.15 (d, J=8.2, 2H), 7.25-7.46 (m, 6H), 7.65 (d,J=8.2, 2H), 7.75 (br s, 1H), 7.84 (d, J=8.6, 1H), 10.71 (s, 1H).

Example 70 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzoyl-L-phenylalanyl)-L-lysinemethyl ester

To a stirred solution ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine methyl ester (369 mg,1 mmol, example 65, step C) in THF/K₂CO₃(1M) (3 mL/3 mL) was addedNα-benzoyl-L-phenylalanine N-hydroxysuccinimide ester (440 mg, 1.2mmol). The reaction mixture was stirred overnight, then diluted with 1NHCl and extracted with EtOAc. The organic layer was dried (MgSO₄) andconcentrated. The crude was purified by flash chromatography usinghexane/EtOAc as eluent to afford the desired product (82% yield).

¹H NMR (DMSO-d₆): δ 0.83 (d, J=6.3, 3H), 0.85 (d, J=6.8, 3H), 1.22-1.50(m, 4H), 1.55-1.70 (m, 1H), 1.80-2.00 (m, 2H), 2.43 (s, 3H), 2.90 and3.00 (ABX, J=14.0, 7.5, 2H), 3.12-3.30 (m, 4H), 3.50 (s, 3H), 4.39 (m,1H), 4.82 (m, 1H), 5.95 (br s, 1H), 6.98 (br s, 1H), 7.20-7.52 (m, 10H),7.65-7.75 (m, 4H).

Example 71 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-thiophenylalanyl]-L-lysinemethyl ester

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-phenylalanyl]-L-lysinemethyl ester, product of example 68, following the indications ofgeneral procedure F. The thioamide was obtained in 64% yield.

¹H NMR (DMSO-d₆): δ 0.83 (d, J=6.8, 3H), 0.86 (d, J=6.8, 3H), 1.30 (m,2H), 1.48-1.60 (m, 2H), 1.70 (m, 1H), 1.80-1.95 (m, 2H), 2.42 (s, 3H),2.46 (s, 3H), 2.88 and 3.20 (ABX, J=13.5, 7.5, 2H), 3.00 (m, 2H), 3.45(m, 2H), 3.52 (s, 3H), 4.18 (m, 1H), 4.40 (m, 1H), 5.37 (d, J=7.0, 1H),7.00 (d, J=8.0, 2H), 7.18 (m, 5H), 7.32 (d, J=7.5, 2H), 7.50 (d, J=7.5,2H), 7.75 (d, J=8.0, 2H), 7.97 (br s, 11H).

Example 72 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-thiophenylalanyl]-L-lysine

Saponification ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-thiophenylalanyl]-L-lysinemethyl ester (example 71) using the conditions described in example 65(step B) yielded 87% of the desired material.

¹H NMR (DMSO-d₆): δ 0.80 (d, J=6.5, 3H), 0.83 (d, J=6.8, 3H), 1.00-1.42(m, 5H), 1.78 (m, 1H), 1.90 (m, 1H), 2.33 (s, 3H), 2.37 (s, 3H), 2.75(m, 1H), 2.82-3.15 9m, 6H), 4.15 (t, J=7.2, 1H), 4.30 (m, 1H), 7.00-7.25(m, 7H), 7.38 (d, J=8.2, 2H), 7.50 (d, J=8.3, 2H), 7.68 (d, J=8.2, 2H),7.75 (d, J=9.0, 1H), 9.77 (s, 1H), 12.70 (br s, 1H).

Example 73 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-thiotryptophanyl]-L-lysine

Saponification ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-thiotryptophanyl]-L-lysinemethyl ester (example 75) using the conditions described in example 65(step B) yielded 88% of the desired material.

¹H NMR (DMSO)-d₆): δ 0.80 (d, J=6.5, 3H), 0.83 (d, J=6.8, 3H), 1.00-1.42(m, 5H), 1.73 (m, 1H), 1.83 (m, 1H), 2.28 (s, 3H), 2.37 (s, 3H),2.80-3.02 (m, 3H), 3.03-3.15 (m, 3H), 4.15 (t, J=6.5, 1H), 4.30 (m, 1H),6.90 (t, J=7.4, 1H), 7.00 (t, J=7.4, 1H), 7.10 (s, 1H), 7.20 (d, J=8.0,2H), 7.28 (d, J=8.2, 1H), 7.30-7.42 (m, 4H), 7.67 (m, 4H), 9.68 (s, 1H),10.72 (s, 1H), 12.76 (br s, 1H).

Example 74 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzenesulfonyl-L-thiotryptophanyl)-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzenesulfonyl-L-tryptophanyl)-L-lysine,product of example 4, following the indications of general procedure F.The thioamide was obtained in 34% yield.

¹H NMR (DMSO-d₆): δ 0.80 (d, J=6.5, 3H), 0.83 (d, J=6.8, 3H), 1.00-1.42(m, 5H), 1.73 (m, 1H), 1.83 (m, 1H), 2.37 (s, 3H), 2.85-3.02 (m, 3H),3.03-3.15 (m, 3H), 4.15 (t, J=6.5, 1H), 4.30 (m, 1H), 6.90 (t, J=7.4,1H), 7.00 (t, J=7.4, 1H), 7.10 (s, 1H), 7.27 (d, J=8.0, 2H), 7.32-7.50(m, 5H), 7.55 (d, J=8.5, 2H), 7.67 (d, J=8.5, 2H), 7.81 (d, J=8.5, 2H),9.69 (s, 1H), 10.72 (s, 1H), 12.76 (br s, 1H).

Example 75 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-thiotryptophanyl]-L-lysinemethyl ester

The title compound was prepared fromN-isobutyl-N-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-tryptophanyl]-L-lysinemethyl ester, product of example 69, following the indications ofgeneral procedure F. The thioamide was obtained in 55% yield.

¹H NMR (CDCl₃): δ 0.83 (d, J=6.8, 3H), 0.87 (d, J=6.8, 3H), 1.20-1.62(m, 5H), 1.80-1.98 (m, 2H), 2.33 (s, 3H), 2.44 (s, 3H), 2.88 and 3.02(ABX, J=13.5, 7.5, 2H), 3.22 and 3.33 (ABX, J=14.0, 7.5, 2H), 3.40-3.52(m, 2H), 3.49 (s, 3H), 4.25 (m, 1H), 4.39 (m, 1H), 5.30 (s, 1H), 5.40(d, J=6.0, 1H), 7.00-7.20 (m, 5H), 7.25-7.50 (m, 5H), 5.70 (d, J=7.5,2H), 7.90 (t, J=5.0, 1H), 8.48 (s, 1H).

Example 76 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-thiobenzoyl-L-thiophenylalanyl)-L-lysinemethyl ester

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzoyl-L-phenylalanyl)-L-lysinemethyl ester, product of example 70, following the indications ofgeneral procedure F. The thioamide was obtained in 81% yield.

¹H NMR (DMSO-d₆): δ 0.83 (d, J=6.3, 3H), 0.86 (d, J=6.8, 3H), 1.12-1.50(m, 4H), 1.55-1.70 (m, 1H), 1.80-2.00 (m, 2H), 2.45 (s, 3H), 2.87 (dd,J=14.0, 7.5, 2H), 2.98-3.10 (m, 2H), 3.40-3.50 (m, 2H), 3.51 (s, 3H),3.70 (m, 1H), 4.39 (m, 1H), 5.50 (br s, 1H), 7.20-7.50 (m, 10H),7.65-7.75 (m, 4H), 9.15 (br s, 1H).

Example 77 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-thiobenzoyl-L-thiophenylalanyl)-L-lysine

Saponification ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-thiobenzoyl-L-thiophenylalanyl)-L-lysinemethyl ester (example 76) using the conditions described in example 65(step E) yielded 80% of the desired material.

¹H NMR (DMSO-d₆): δ 0.81 (t, J=6.5, 6H), 1.15-1.52 (m, 5H), 1.80-1.90(m, 2H), 2.36 (s, 3H), 2.92 and 2.98 (ABX, J=14.5, 7.5, 2H), 3.10 (m,1H), 3.30-3.50 (m, 3H), 4.20 (t, J=7.0, 1H), 5.60 (m, 1H), 7.18-7.28 (m,3H), 7.32-7.50 (m, 5H), 7.68 (m, 4H), 10.23 (m, 2H), 12.80 (br s, 1H).

Example 78 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-L-phenylalanyl)-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine hydrochloride (1.2 g,3.1 mmol, example 1, step E) suspended in THF (50 mL) and 1N NaOH (0.5mL, to reach pH 10) as described in general procedure Bc usingcommercially available Nα-benzyloxycarbonyl-L-phenylalanine (1.0 g, 3.5mmol), N-hydroxysuccinimide (0.4 g, 3.5 mmol) and DCC (1.1 g, 4.8 mmol).The final product was triturated with ether to yield 1.61 g (95%) of thedesired material.

¹H NMR (CDCl₃): δ 0.83 (m, 6H), 1.08-1.11 (m, 2H), 1.23-1.25 (m, 2H),1.35 (s, 9H), 1.45-1.52 (m, 1H), 1.89-1.99 (m, 2H), 2.36 (s, 3H),2.94-3.09 (m, 6H), 4.20-4.23 (m, 2H), 7.09-7.26 (m, 7H), 7.73 (d, J=8.1,2H). LC-MS: 602.2 (M−H)⁻, 95% pure.

Example 79 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(L-phenylalanyl)-L-lysine

This product was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-L-phenylalanyl)-L-lysine(600 mg, 1.0 mmol, example 78) as described in general procedure C. Theresidue was triturated with ether and placed under high vacuum to yielda hard white foam (440 mg, 88%).

¹H NMR (CDCl₃): δ0.78 (d, J=6.3, 3H), 0.81 (d, J=6.3, 3H), 1.00-1.11 (m,2H), 1.23-1.25 (m, 2H), 1.44-1.47 (m, 1H), 1.70-1.74 (m, 1H), 1.89-1.93(m, 1H), 2.32 (s, 3H), 2.82-2.89 (m, 2H), 2.91-2.95 (m, 2H), 3.88 (br s,1H), 4.17 (t, J=5.9, 1H), 7.14 (d, J=8.0, 2H), 7.23 (t, J=4.9, 1H), 7.29(t, J=4.8, 2H), 7.31 (d, J=5.0, 2H), 7.64 (d, J=8.1, 2H), 8.05 (br s,3H). LC-MS: 502.2 (M−H)⁻, 95% pure.

Example 80 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(carbotetrahydro-3-furanyloxy)-L-phenylalanyl]-L-lysine

Step A. Preparation of Nα-(carbotetrahydro-3-furanyloxy)-L-phenylalanine

L-phenylalanine was reacted with tetrahydro-3-furanyloxy-1-nitrophenylcarbonate under the conditions used in general procedure A giving thetitle compound which was recrystallised from DCM (28%). This compoundwas used without further purification in the next step.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(carbotetrahydro-3-furanyloxy)-L-phenylalanyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingNα-(carbotetrahydro-3-furanyloxy)-L-phenylalanine (84 mg, 0.3 mmol)which was prepared in step A of this example. The final product wastriturated with ether to yield 51 mg (27%) of the desired material.

LC-MS: 618.9 (M+H)⁺, 90% pure.

Example 81 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-N′α-methyl-L-phenylalanyl)-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingcommercially available Nα-tert-butoxycarbonyl-Nα-methyl-L-phenylalanine(83 mg, 0.3 mmol). The final product was triturated with ether to yield176 mg (90%) of the desired material.

LC-MS: 616.8 (M−H)⁻, 94% pure.

Example 82 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-L-methionyl)-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingcommercially available Nα-tert-butoxycarbonyl-L-methionine (75 mg, 0.3mmol). The final product was triturated with ether to yield 170 mg (96%)of the desired material.

LC-MS: 586.8 (M−H)⁻, 90% pure.

Example 83 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-tert-butoxycarbonyl-S-(4-methylbenzyl)-L-cysteinyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingcommercially availableNα-tert-butoxycarbonyl-S-(4-methylbenzyl)-L-cysteine (67 mg, 0.3 mmol).The final product was triturated with ether to yield 130 mg (65%) of thedesired material.

LC-MS: 562.8 (M−H)⁻, 95% pure.

Example 84 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-O-benzyl-L-threonyl)-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingcommercially available Nα-tert-butoxycarbonyl-O-benzyl-L-threonine (93mg, 0.3 mmol). The final product was triturated with ether to yield 195mg (98%) of the desired material.

LC-MS: 646.9 (M−H)⁻, 95% pure.

Example 85 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-Nτ-benzyl-L-histidinyl)-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingcommercially available Nα-tert-butoxycarbonyl-Nτ-benzyl-L-histidine (100mg, 0.3 mmol). The final product was triturated with ether to yield 133mg (65%) of the desired material.

LC-MS: 682.2 (M−H)⁻, 90% pure.

Example 86 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-L-tryptophanyl)-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingcommercially available Nα-tert-butoxycarbonyl-L-tryptophan (90 mg, 0.3mmol). The final product was triturated with ether to yield 157 mg (81%)of the desired material.

LC-MS: 641.8 (M−H)⁻, 90% pure.

Example 87 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-O-benzyl-L-tyrosyl)-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol,example 1, step E) as described in general procedure Bc usingcommercially available Nα-tert-butoxycarbonyl-O-benzyl-L-tyrosine (111mg, 0.3 mmol). The final product was triturated with ether to yield 145mg (68%) of the desired material.

LC-MS: 708.2 (M−H)⁻, 95% pure.

Example 88 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-methyl-L-phenylalanyl)-L-lysine

This product was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-N′α-methyl-L-phenylalanyl)-L-lysine(100 mg, 0.16 mmol, example 81) as described in general procedure C. Theresidue was triturated with ether and placed under high vacuum to yielda hard white foam (25 mg, 30%).

LC-MS: 516.3 (M−H)⁻, 95% pure.

Example 89 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(L-methionyl)-L-lysine

This product was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-L-methionyl)-L-lysine(95 mg, 0.16 mmol, example 82) as described in general procedure C. Theresidue was triturated with ether and placed under high vacuum to yielda hard white foam (40 mg, 51%).

LC-MS: 486.2 (M−H)⁻, 95% pure.

Example 90 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[S-(4-methylbenzyl)-L-cysteinyl]-L-lysine

This product was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-tert-butoxycarbonyl-S-(4-methylbenzyl)-L-cysteinyl]-L-lysine(91 mg, 0.16 mmol, example 83) as described in general procedure C. Theresidue was triturated with ether and placed under high vacuum to yielda hard white foam (60 mg, 66%).

LC-MS: 562.2 (M−H)⁻, 85% pure.

Example 91 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(O-benzyl-L-threonyl)-L-lysine

This product was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-O-benzyl-L-threonyl)-L-lysine(110 mg, 0.18 mmol, example 84) as described in general procedure C. Theresidue was triturated with ether and placed under high vacuum to yielda hard white foam (40 mg, 41%).

LC-MS: 549.3 (M−H)⁻, 95% pure.

Example 92 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(Nτ-benzyl-L-histidinyl)-L-lysine

This product was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-Nτ-benzyl-L-histidinyl)-L-lysine(110 mg, 0.16 mmol, example 85) as described in general procedure C. Theresidue was triturated with ether and placed under high vacuum to yielda hard white foam (51 mg, 54%).

LC-MS: 582.1 (M−H)⁻, 95% pure.

Example 93 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(L-tryptophanyl)-L-lysine

This product was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-L-tryptophanyl)-L-lysine(100 mg, 0.15 mmol, example 86) as described in general procedure C. Theresidue was triturated with ether and placed under high vacuum to yielda hard white foam (27 mg, 33%).

LC-MS: 541.6 (M−H)⁻, 90% pure.

Example 94 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(O-benzyl-L-tyrosyl)-L-lysine

This product was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-O-benzyl-L-tyrosyl)-L-lysine(115 mg, 0.16 mmol, example 87) as described in general procedure C. Theresidue was triturated with ether and placed under high vacuum to yielda hard white foam (19 mg, 19%).

LC-MS: 608.2 (M−H)⁻, 90% pure.

Example 95 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-benzoyl-S-phenylalanyl)-2,6-diaminohexanol

The product of example 70,Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzoyl-L-phenylalanyl)-L-lysinemethyl ester, was reduced with LiAlH₄ following the indications ofgeneral procedure G. The final product was obtained in 76% yield.

¹H NMR (DMSO-d₆): δ0.77 (t, J=7.0, 6H), 0.90-1.30 (m, 5H), 1.48 (m, 1H),1.85 (m, 1H), 2.38 (s, 3H), 2.55-2.70 (m, 3H), 2.75-2.85 (m, 2H), 2.93(dd, J=13.5, 7.5, 1H), 3.35 (m, 1H), 3.50 (m, 1H), 3.86 (m, 1H), 4.65(m, 1H), 7.10 (d, J=7.8, 2H), 7.12-7.22 (m, 6H), 7.35 (d, J=8.0, 2H),7.50 (d, J=7.8, 2H), 7.67 (d, J=8.2, 2H), 7.72(t, J=5.0, 1H), 7.70 (d,J=8.0, 1H).

Example 96 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-(4-methylbenzenesulfonyl)-S-phenylalanyl]-2,6-diaminohexanol

The product of example 68,Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-phenylalanyl]-L-lysinemethyl ester, was reduced with LiAlH₄ following the indications ofgeneral procedure G. The final product was obtained in 76% yield.

¹H NMR (DMSO-d₆): δ0.83 (t, J=7.0, 6H), 0.90-1.30 (m, 5H), 1.52 (m, 1H),1.90 (m, 1H), 2.36 (s, 3H), 2.80 (dd, J=12.0, 8.0, 1H), 2.85-3.10 (m,5H), 3.52 (m, 1H), 4.66 (m, 2H), 7.10-7.28 (m, 3 H), 7.30-7.55 (m, 7H),7.67 (d, J=8.0, 2H), 7.80 (d, J=8.0, 2H), 7.96 (m, 1H), 8.51 (d, J=8.0,1H).

Example 97 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-(4-methylbenzenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanol

The product of example 69,Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-tryptophanyl]-L-lysinemethyl ester, was reduced with LiAlH₄ following the indications ofgeneral procedure G. The final product was obtained in 65% yield.

¹H NMR (DMSO-d₆): δ0.82 (d, J=7.0, 3H), 0.85 (d, J=6.8, 3H), 0.88-1.20(m, 5H), 1.45 (m, 1H), 2.30 (s, 3H), 2.36 (s, 3H), 2.62 (m, 2H), 2.76(m, 2H), 2.90 (m, 2H), 2.90 (m, 2H), 3.20-3.40 (m, 2H), 3.50 (m, 1H),3.85 (m, 1H), 4.67 (t, J=5.0, 1H), 6.90 (t, J=7.4, 1H), 7.03 (m, 2H).7.13 (d, J=7.6, 2H), 7.27 (d, J=7.6, 2H), 7.35 (m, 3H), 7.46 (d, J=7.6,2H), 7.68 (m, 3H), 7.82 (d, J=8.8, 1H), 10.70 (s, 1H).

Example 98 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-phenylalanyl-N-cyanoamidine]-L-lysine

To a stirred solution of the thioamide,Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-thiophenylalanyl]-L-lysine,(227 mg, 0.33 mmol, example 72) in MeOH (3 mL) was added cyanamide (28mg, 0.66 mmol). The mixture was stirred for 5 min, then mercuric acetate(209 mg, 0.66 mmol) was added. The reaction was stirred for 3 h thendiluted with saturated NH₄Cl and extracted with EtOAc. The organic layerwas washed with brine and concentrated then rediluted with THF/MeOH (2mL/1 mL) and treated with 1N NaOH (0.8 mL). After stirring for 4 h, thereaction was acidified with 1N HCl and extracted with EtOAc. The organiclayer was dried over MgSO₄, concentrated and purified by columnchromatography to give 128 mg (57%) of the final product.

¹H NMR (DMSO-d₆): δ0.80 (d, J=6.5, 3H), 0.83 (d, J=6.8, 3H), 1.00-1.12(m, 4H), 1.35 (m, 1H), 1.71 (m, 1H), 1.88 (m, 1H), 2.37 (s, 3H), 2.50(s, 3H), 2.60-2.80 (m, 4H), 2.90 and 2.97 (ABX, J=13.2, 7.0, 2H), 3.88(m, 1H), 4.15 (t, J=7.0, 1H), 7.10 (d, J=8.2, 2H), 7.13-7.23 (m, 5H),7.38 (d, J=8.2, 2H), 7.49 (d, J=7.9, 2H), 7.68 (d, J=8.2, 2H), 7.78 (m,1H), 7.92 (d, J=8.3, 1H), 12.70 (br s, 1H).

Example 99 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-acetyl-L-tryptophanyl)-L-lysine

Step A. Preparation of Nα-acetyl-L-tryptophan

L-tryptophan was reacted with acetyl chloride under the conditions usedin general procedure A giving the title compound which wasrecrystallised from DCM (65%).

¹H NMR (CDCl₃): δ 1.92 (s, 3H), 3.11-3.46 (m, 2H), 4.73 (t, J=4.5, 1H),6.92-7.10 (m, 3H), 7.28 (d, J=6.0, 1H), 7.55 (d, J=6.0, 1H).

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-acetyl-L-tryptophanyl)-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine hydrochloride (400 mg,1.0 mmol, example 1, step E) suspended in THF (20 mL) and 1N NaOH (0.5mL, to reach pH 10) as described in general procedure Bc usingNα-acetyl-L-tryptophan (300 mg, 1.4 mmol) prepared in step A of thisexample, N-hydroxysuccinimide (115 mg, 1.0 mmol) and DCC (210 mg, 1.0mmol). The crude material (600 mg) was purified by preparative HPLC toyield 411 mg (45%) of the desired material.

¹H NMR (CDCl₃): δ0.82 (d, J=6.8, 3H), 0.84 (d, J=6.8, 3H), 1.08-1.11 (m,2H), 1.18-1.22 (m, 2H), 1.45-1.52 (m, 1H), 1.94 (s, 3H), 2.32 (s, 3H),2.84-3.09 (m, 4H), 3.15-3.18 (m, 1H), 4.19 (t, J=7.5, 1H), 4.57 (t,J=7.1, 1H), 6.85 (t, J=7.2, 1H), 7.04 (s, 2H), 7.32 (d, J=8.1, 2H), 7.56(d, J=8.1, 1H), 7.69 (d, J=7.2, 2H). LC-MS: 583.8 (M−H)⁻, 99% pure.

Example 100 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-pivaloyl-L-tryptophanyl)-L-lysine

Step A. Preparation of Nαpivaloyl-L-tryptophan

L-tryptophan was reacted with pivaloyl chloride under the conditionsused in general procedure A giving the title compound which wasrecrystallised from DCM (50%).

¹H NMR (CDCl₃): δ1.02 (s, 9H), 3.21-3.46 (m, 2H), 4.73 (t, J=4.5, 1H),6.92-7.10 (m, 3H), 7.28 (d, J=6.0, 1H), 7.55 (d, J=6.0, 1H).

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-pivaloyl-L-tryptophanyl)-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine hydrochloride (400 mg,1.0 mmol, example 1, step E) suspended in THF (20 mL) and 1N NaOH (0.5mL, to reach pH 10) as described in general procedure Bc usingNα-pivaloyl-L-tryptophan (350 mg, 1.5 mmol) prepared in step A of thisexample, N-hydroxysuccinimide (115 mg, 1.0 mmol) and DCC (210 mg, 1.0mmol). The crude material (488 mg) was purified by preparative HPLC toyield 244 mg (50%) of the desired material.

¹H NMR (CDCl₃): δ0.82(d, J=6.8, 3H), 0.84 (d, J=6.8, 3H), 1.06 (s, 9H),1.08-1.11 (m, 2H), 1.18-1.22 (m, 2H), 1.45-1.52 (m, 1H), 2.38 (s, 3H),2.84-3.09 (m, 4H), 3.15-3.18 (m, 1H), 4.24 (t, J=7.5, 1H), 4.57 (t,J=7.1, 1H), 6.85 (t, J=7.2, 1H), 7.04 (s, 2H), 7.32 (d, J=8.1, 2H), 7.56(d, J=8.1, 1H), 7.69 (d, J=7.2, 2H). LC-MS: 625.8 (M−H)⁻, 99% pure.

Example 101 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-trifluoroacetyl-L-phenylalanyl)-L-ysine

Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-L-phenylalanyl)-L-lysine(50 mg, 0.08 mmol, example 78) is added to a mixture of oftrifluoroacetic anhydride and TFA and was stirred for a period of 30min. Afterwards, the reactant were evaporated and the residue trituratedwith ether to give 40 mg (74%) of the title compound.

LC-MS: 598.7 (M−H)⁻, 99% pure.

Example 102 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-(4-methylbenzenesulfonyl)-S-thiotryptophanyl]-2,6-diaminohexanol

The product of example 75,Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-thiotryptophanyl]-L-lysinemethyl ester, was reduced with LiAlH₄ following the indications ofgeneral procedure G. The final product was obtained in 61% yield.

¹H NMR (CDCl₃): δ0.83 (d, J=7.0, 3H), 0.85 (d, J=7.0, 3H), 0.88-1.25 (m,5H), 1.50 (m, 1H), 1.88 (m, 1H), 2.30 (s, 3H), 2.37 (s, 3H), 2.80 and2.95 (ABX, J=14.5, 7.3, 2H), 2.90 and 3.10 (ABX, J=14.5, 7.8, 2H), 3.05(m, 2H), 3.20-3.40 (m, 2H), 3.51 (m, 1H), 4.30 (m, 1H), 4.86 (m, 1H),6.90 (t, J=7.5, 1H), 7.02 (t, J=7.5, 1H), 7.06 (s, 1H), 7.09 (d, J=8.0,2H), 7.28 (d, J=7.7, 1H), 7.32-7.42 (m, 5H), 7.70 (d, J=8.10, 4H), 10.71(s, 1H).

Example 103 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-tryptophanyl-N-cyanoamidine]-L-lysine

To a stirred solution of thioamideNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-thiotryptophanyl]-L-lysine(240 mg, 0.33 mmol, example 73) in MeOH (3 mL) was added cyanamide (28mg, 0.66 mmol). The mixture was stirred for 5 min, then mercuric acetate(209 mg, 0.66 mmol) was added. The reaction was stirred for 3 h thendiluted with saturated NH₄Cl and extracted with EtOAc. The organic layerwas washed with brine and concentrated then rediluted with THF/MeOH (2mL/1 mL) and treated with 1N NaOH (0.8 mL). After stirring for 4 h, thereaction was acidified with 1N HCl and extracted with EtOAc. The organiclayer was dried over MgSO₄, concentrated and purified by columnchromatography to give 154 mg (65%) of the final product.

¹H NMR (DMSO-d₆): δ0.80 (d, J=6.5, 3H), 0.83 (d, J=6.8, 3H), 1.00-1.14(m, 4H), 1.38 (m, 1H), 1.72 (m, 1H), 1.88 (m, 1H), 2.30 (s, 3H), 2.37(s, 3H), 2.70 (dd, J=13.5, 7.0, 1H), 2.92 (m, 3H), 3.87 (m, 1H), 4.12(t, J=7.0, 1H), 6.90 (t, J=7.5, 1H), 7.00 (t, J=7.5, 1H), 7.04 (s, 1H),7.13 (d, J=8.3, 2H), 7.28 (d, J=8.3, 1H), 7.38 (m, 3H), 7.47 (d, J=8.0,2H), 7.65 (d, J=8.0, 2H), 7.72 (t, J=5.0, 1H), 7.85 (d, J=8.1, 1H),10.73 (s, 1H), 12.70 (br s, 1H).

Example 104 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzenesulfonyl-L-tryptophanyl)-L-lysine2,3-dihydroxypropyl ester

Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzenesulfonyl-L-tryptophanyl)-L-lysine(170 mg, 0.25 mmol) dissolved in a solution of DMF (1 mL) was treatedwith glycerol (100 μL, 1.37 mmol) and EDC (100 mg, 0.5 mmol) and stirredovernight. Then, the solution was poured in 5% citric acid and extractedwith EtOAc. The solvent was evaporated and the residue purified bypreparative HPLC to give 150 mg (79%) of the final product.

¹H NMR (CDCl₃): δ0.81 (d, J=6.3, 3H), 0.83 (d, J=6.3, 3H), 0.94-1.03 (m,4H), 1.32-1.35 (m, 1H), 1.46-1.49 (m, 1H), 1.81-1.93 (m, 2H), 2.35 (s,3H), 2.75 (br s, 1H), 2.82-2.99 (m, 4H), 3.45 (br s, 2H), 3.65 (br s,1H), 3.82 (br s, 2H), 4.21 (t, J=7.2, 1H), 6.85 (t, J=4.5, 1H), 7.00 (t,J=4.5, 1H), 7.23-7.31 (m, 6H), 7.42 (t, J=4.5, 1H), 7.60 (d, J=6.8, 2H),7.73 (d, J=6.8, 2H).

Example 105 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-phenylglycyl)-L-lysine

Step A. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-chloroacetyl-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine potassium salt (2.0 g,5.1 mmol) suspended in a mixture of THF (20 mL), DIEA (1.5 mL, 8.6 mmol)and chloroacetyl chloride (0.8 mL, 10.1 mmol). The reaction mixture wasstirred for a period of 30 min. Then, a solution of 2N HCl (20 mL) wasadded and the solution extracted with EtOAc (25 mL, 3×). The organicphase was dried with MgSO₄, filtered and evaporated to a brown oil. Thecrude material was purified by flash chromatography using a solventgradient from 39:1 to 19:1 CH₂Cl₂/MeOH. The product was isolated as ayellowish oil (1.7 g, 78% yield).

Rf=0.43 (EtOAc/hexane, 9:1). LC-MS: 464 (M+H)⁺, 90% pure.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-iodoacetyl-L-lysine

To a solution ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-chloroacetyl-L-lysine (1.7g, 3.9 mmol) in dry acetone (20 mL) was added NaI (1.5 g, 10.0 mmol).The resulting mixture was stirred at room temperature for a period of 24h. The precipitate NaCl was filtered off and the solvent evaporated toan oil. The crude material was dissolved in CH₂Cl₂ to precipitate theremaining NaI which was filtered off. The filtrate was washed with a 5%aqueous solution of NaHSO₃ (10 mL) and with water (25 mL, 3×). Theorganic solvent was dried, filtered and evaporated to a brown oil (1.98g, 96% yield) which was used without further purification in the nextstep.

¹H NMR (DMSO-d₆): δ0.81 (d, J=6.5, 3H), 0.83 (d, J=6.3, 3H), 1.20 (m,2H), 1.33 (m, 2H), 1.45 (m, 1H), 1.81 (m, 1H), 1.89 (m, 1H), 2.38 (s,3H), 2.95 (m, 4H), 3.60 (s, 2H), 4.18 (t, J=7.2, 1H), 7.38 (d, J=7.6,2H), 7.67 (d, J=7.6, 2H), 8.17 (t, J=5.0, 1H), 12.70 (br s, 1H). LC-MS:556 (M+H)⁺, 99% pure.

Step C. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-phenylglycyl)-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-iodoacetyl-L-lysine (200 mg,0.38 mmol, product of step B above) by following the indications ofgeneral procedure H using DIEA (0.19 mL, 1.09 mmol) and aniline (0.16mL, 1.76 mmol). The crude material was purified by preparative HPLC. Theproduct was isolated as a solid (90 mg, 48% yield).

¹H NMR (DMSO-d): δ0.79 (d, J=6.7, 3H), 0.81 (d, J=6.9, 3H), 1.15 (m,2H), 1.32 (m, 2H), 1.41 (m, 1H), 1.77 (m, 1H), 1.88 (m, 1H), 2.37 (s,3H), 2.90-3.01 (m, 4H), 3.58 (d, J=3.1, 2H), 4.14 (t, J=7.1, 1H), 5.86(s, 1H), 6.52 (d, J=7.7, 2H), 6.57 (t, J=7.1, 1H), 7.08 (t, J=7.6, 2H),7.36 (d, J=7.6, 2H), 7.66 (d, J=8.2, 2H), 7.80 (t, J=5.0, 1H), 12.5 (s,1H). LC-MS: 490 (M+H)⁺, 99% pure.

Example 106 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(3-pyridyl)glycyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-iodoacetyl-L-lysine (200 mg,0.38 mmol, example 105, step B) by following the indications of generalprocedure H using DIEA (0.19 mL, 1.09 mmol) and 3-aminopyridine (175 mg,1.36 mmol). The crude material was purified by preparative HPLC. Theproduct was isolated as a solid (42 mg, 22% yield).

¹H NMR (DMSO-d₆): δ0.75 (d, J=6.7, 3H), 0.81 (d, J=6.7, 3H), 1.23-1.37(m, 4H), 1.46 (m, 1H), 1.84 (m, 1H), 1.99 (m, 1H), 2.31 (s, 3H), 2.85(m, 1H), 3.07 (m, 3H), 3.95 (t, J=6.6, 1H), 5.21 (d, J=9.3, 2H), 7.01(s, 2H), 7.26 (d, J=8.0, 2H), 7.55 (d, J=8.6, 2H), 7.62 (t, J=7.0, 1H),7.75 (d, J=7.8, 2H), 8.03 (d, J=5.4, 1H), 8.21 (s, 1H), 9.28 (t, J=4.8,1H). LC-MS: 491 (M+H)⁺, 95% pure.

Example 107 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(2,3-dimethoxybenzyl)glycyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-iodoacetyl-L-lysine (150 mg,0.29 mmol, example 105, step B) by following the indications of generalprocedure H using DIEA (0.14 mL, 0.81 mmol) and 2,3-dimethoxybenzylamine(0.12 mL, 0.81 mmol). The crude material was purified by preparativeHPLC. The product was isolated as a solid (25 mg, 16% yield).

¹H NMR (DMSO-d₆): δ0.79 (d, J=6.9, 3H), 0.81 (d, J=6.9, 3H), 1.20 (m,2H), 1.30-1.5 (m, 3H), 1.75-1.95 (m, 2H), 2.36 (s, 3H), 2.90-3.05 (m,4H), 3.09 (s, 2H), 3.68 (s, 2H), 3.72 (s, 3H), 3.79 (s, 3H), 4.16 (t,J=6.6, 1H), 6.95 (m, 2H), 7.01 (d, J=7.6, 1H), 7.35 (d, J=7.9, 2H), 7.67(d, J=8.0, 2H), 7.80 (t, J=5.4, 1H). LC-MS: 564 (M+H)⁺, 98% pure.

Example 108 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(2-pyridyl)glycyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-iodoacetyl-L-lysine (200 mg,0.38 mmol, example 105, step B) by following the indications of generalprocedure H using DIEA (0.19 mL, 1.09 mmol) and 2-aminopyridine (170 mg,1.31 mmol). The crude material was purified by preparative HPLC. Theproduct was isolated as a solid (88 mg, 47% yield).

¹H NMR (DMSO-d₆): δ0.79 (d, J=6.1, 3H), 0.82 (d, J=6.4, 3H), 1.26 (m,2H), 1.39-1.50 (m, 3H), 1.75-1.90 (m, 2H), 2.37 (s, 3H), 2.87-3.04 (m,4H), 4.21 (t, J=6.6, 1H), 4.90 (s, 2H), 6.88 (t, J=6.5, 1H), 7.05 (d,J=8.9, 1H), 7.37 (d, J=7.9, 2H), 7.67 (d, J=8.1, 2H), 7.88 (t, J=8.1,1H), 7.95 (d, J=6.3, 1H,), 8.33 (t, J=5.6 1H), 8.41 (s, 2H), 12.7 (s,1H). LC-MS: 491 (M+H)⁺, 99% pure.

Example 109 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzylglycyl)-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-iodoacetyl-L-lysine (200 mg,0.38 mmol, example 105, step B) by following the indications of generalprocedure H using DIEA (0.20 mL, 1.10 mmol) and benzylamine (320 mg,2.99 mmol). The crude material was purified by preparative HPLC. Theproduct was isolated as a solid (137 mg, 71% yield).

¹H NMR (CDCl₃): δ0.81 (s, 6H), 1.26-1.60 (m, 5H), 1.80-2.05 (m, 2H),2.40 (s, 3H), 2.90-3.00 (m, 3H), 3.04 (s, 1H), 3.19 (m, 2H), 3.87 (s,1H), 4.26 (s, 2H), 6.16 (s, 4H), 7.28 (d, J=5.3, 2H), 7.38 (m, 3H), 7.48(d, J=4.5, 2H), 7.70 (d, J=7.5, 2H), 8.90 (s, 1H). LC-MS: 502 (M−H)⁻,99% pure.

Example 110 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(piperidinyl-N-ethyl)glycyl]-L-lysinetrifluoroacetic acid salt

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-iodoacetyl-L-lysine (150 mg,0.29 mmol, example 105, step B) by following the indications of generalprocedure H using DIEA (0.12 mL, 0.69 mmol) and1-(2-aminoethyl)piperidine (0.12 mL, 0.84 mmol). The crude material waspurified by preparative HPLC. The product was isolated as a solid (65mg, 33% yield).

¹H NMR (DMSO-d₆): δ0.80 (d, J=6.3, 3H), 0.82 (d, J=6.7, 3H), 1.24-2.00(m, 13H), 2.38 (s, 3H), 2.85-3.40 (m, 12H), 3.76 (s, 2H), 4.21 (t,J=6.7, 1H), 7.37 (d, J=7.9, 2H), 7.67 (d, J=8.0, 2H), 8.49 (t, J≅5.0,1H), 9.25 (s, 2H). LC-MS: 525 (M+H)⁺, 99% pure.

Example 111 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-morpholinyl-N-ethyl)glycyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-iodoacetyl-L-lysine (150 mg,0.29 mmol, example 105, step B) by following the indications of generalprocedure H using DIEA (0.12 mL, 0.69 mmol) and4-(2-aminoethyl)morpholine (0.11 mL, 0.84 mmol). The crude material waspurified by preparative HPLC. The product was isolated as a solid (51mg, 34% yield).

¹H NMR (DMSO-d₆): δ0.80 (d, J=6.1, 3H), 0.82 (d, J=7.0, 3H), 1.24-1.30(m, 2H), 1.35-1.55 (m, 3H), 1.75-1.95 (m, 2H), 2.38 (s, 3H), 2.80-3.25(m, 10H), 3.30 (s, 2H), 3.75 (s, 6H), 4.21 (t, J=7.1, 1H), 7.37 (d,J=8.3, 2H), 7.68 (d, J=8.4, 2H), 8.47 (m, 1H). LC-MS: 525 (M−H)⁻, 99%pure.

Example 112 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-pyridyl)glycyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-iodoacetyl-L-lysine (150 mg,0.29 mmol, example 105, step B) by following the indications of generalprocedure H using DIEA (0.10 mL, 0.57 mmol) and 4-aminopyridine (76 mg,0.59 mmol). The crude material was purified by preparative HPLC. Theproduct was isolated as a solid (39 mg, 28% yield).

¹H NMR (DMSO-d₆): δ0.79 (d, J=6.8, 3H), 0.81 (d, J=6.5, 3H), 1.25 (m,2H), 1.35-1.51 (m, 3H), 1.78-1.91 (m, 2H), 2.37 (s, 3H), 2.90 (m, 1H),3.00 (m, 3H), 4.20 (t, J=7.1, 1H), 4.85 (s, 2H), 6.80 (d, J=6.7, 2H),7.37 (d, J=8.2, 2H), 7.67 (d, J=7.6, 2H), 8.02 (d, J=7.0, 2H), 8.15 (s,2H), 8.37 (t, J=4.9, 1H), 12.74 (br s, 1H). LC-MS: 489 (M−H)⁻, 99% pure.

Example 113 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(3-quinolyl)glycyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-iodoacetyl-L-lysine (200 mg,0.38 mmol, example 105, step B) by following the indications of generalprocedure H using DIEA (0.19 mL, 1.09 mmol) and 3-aminoquinoline (260mg, 1.80 mmol). The crude material was purified by preparative HPLC. Theproduct was isolated as a solid (24 mg, 12% yield).

¹H NMR (DMSO-d₆): δ0.76 (d, J=5.9, 3H), 0.82 (d, J=6.5, 3H), 1.26-1.50(m, 5H), 1.80-2.10 (m, 2H), 2.34 (s, 3H), 2.86 (m, 1H), 3.04-3.20 (m,2H), 4.00 (m, 1H), 5.67 (d, J=4.8, 2), 6.85 (s, 2H), 7.29 (d, J=7.8,2H), 7.75 (m, 4H), 8.02 (s, 1H), 8.07 (d, J=8.0, 1H), 8.12 (d, J=8.5,1H), 8.97 (m, 1H), 9.05 (s, 1H). LC-MS: 539 (M−H)⁻, 95% pure.

Example 114 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzyl-N′α-phenylglycyl)-L-lysine

Step A. Preparation of N-benzyl-N-phenylglycine

Commercially available N-phenylglycine (1.51 g, 10.0 mmol) dissolved inTHF (20 mL) was treated with butyllithium 2.5M in hexane (8.8 mL, 22.0mmol) under an inert atmophere of argon at −78° C. The mixture wasstirred for 15 min before the addition of benzylbromide (1.3 mL, 11mmol). The reaction mixture was stirred for 2 h while warming up to roomtemperature. Then, 2N HCl (25 mL) was added and the mixture extractedwith EtOAc (30 mL, 3×). The organic phase was dried over MgSO₄, filteredand concentrated to an oil. The crude material was purified by flashchromatography using a solvent gradient from 19:1 to 9:1 CH₂Cl₂/MeOH.The product was isolated as a brown and hardy oil (1.98 g, 82%).

¹H NMR (CDCl₃): δ4.16 (s, 2H), 4.68 (s, 2H), 6.76 (d, J=8.5, 2H), 6.83(t, J=7.2, 1H), 7.27 (t, J=6.1, 2H), 7.33 (m, 3H), 7.38 (t, J=7.6, 2H),10.70 (s, 1H).

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzyl-N′α-phenylglycyl)-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysine(200 mg) in a similar fashion to general procedure Bb usingN-benzyl-N-phenylglycine (100 mg, 0.45 mmol) prepared in step A of thisexample with DCC (200 mg, 0.97 mmol) and HOBt (100 mg, 0.74 mmol) as theactivating reagents. The final product was purified by preparative HPLCto yield 21 mg (75%) of the desired material.

¹H NMR (DMSO-d₆): δ0.79 (d, J=6.9, 3H), 0.82 (d, J=7.2, 3H), 1.17 (m,2H), 1.33 (m, 2H), 1.42 (m, 1H), 1.77 (m, 1H), 1.88 (m, 1H), 2.37 (s,3H), 2.90-3.01 (m, 4H), 3.94 (s, 2H), 4.16 (t, J=7.2, 1H), 4.62 (s, 2H),6.59 (m, 3H), 7.11 (t, J=7.5, 2H), 7.24 (m, 3H), 7.31 (t, J=7.3, 2H),7.36 (d, J=7.6, 2H), 7.66 (d, J=8.5, 2H), 7.87 (t, J=5.3, 1H), 12.6 (s,1H). LC-MS: 578 (M+H)⁺, 85% pure.

Example 115 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-methyl-N′α-phenylglycyl)-L-lysine

Step A. Preparation of N-methyl-N-phenylglycine

The title compound was prepared from N-phenylglycine (200 mg, 1.3 mmol)as described for the preparation of N-benzyl-N-phenylglycine (example114, step A) using methyl iodide (1 mL, 6.6 mmol) instead ofbenzylbromide. The crude material was purified by preparative HPLC. Theproduct was isolated as a solid (90 mg, 41% yield).

¹H NMR (CDCl₃): δ3.07 (s, 3H), 4.09 (s, 2H), 6.73 (d, J=7.7, 2H), 6.80(t, J=7.2, 1H), 7.27 (t, J=7.3, 2H), 8.45 (s, 1H).

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-methyl-N′α-phenylglycyl)-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysine(200 mg) in a similar fashion to general procedure Bb usingN-methyl-N-phenylglycine (75 mg, 0.45 mmol) prepared in step A of thisexample with DCC (200 mg, 0.97 mmol) and HOBt (100 mg, 0.74 mmol) as theactivating reagents. The final product was purified by preparative HPLCto yield 7 mg (25%) of the desired material.

¹H NMR (DMSO-d₆): δ0.79 (d, J=6.5, 3H), 0.82 (d, J=7.0, 3H), 1.16 (m,2H), 1.32 (m, 2H), 1.41 (m, 1H), 1.77 (m, 1H), 1.88 (m, 1H), 2.37 (s,3H), 2.90-3.01 (m, 4H), 2.96 (s, 3H), 3.82 (s, 2H), 4.16 (t, J=7.0, 1H),6.62 (m, 3H), 7.16 (t, J=7.4, 2H), 7.36 (d, J=8.1, 2H), 7.66 (d, J=8.0,2H), 7.80 (t, J=4.3, 1H), 12.7 (s, 1H). LC-MS: 502 (M+H)⁺, 98% pure.

Example 116 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-(4-methylbenzenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanal

Step A. Preparation of (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol

A solution of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (12.0 g,30.5 mmol, example 1, step E) dissolved in MeOH (100 mL) was treatedwith of trimethylsilyl chloride (20 mL). The mixture was refluxed 5 hbefore stirring at room temperature for 5 h. Afterwards, the solutionwas evaporated and placed under high vacuum until a hard foam wasobtained (13.6 g). This was dissolved in dry THF (100 mL) and addeddropwise to a solution of LiAlH₄ (5.0 g, 131.6 mmol) in THF (300 mL).The solution was stirred for 2 h, then heated to reflux for 20 min.After cooling in an ice bath the solution was quenched by addition ofMeOH (5 mL), water (5 mL), then 10% NaOH (5 mL). The solvent wasevaporated and the precipitate dissolved in MeOH (200 mL) was stirredfor 2 h at 60° C. The granular precipitate formed was filtered offthrough celite, and the liquor was concentrated to form a clear oilwhich solidified on standing (8.48 g, 81%).

¹H NMR (CDCl₃): δ0.85 (d, J=6.3, 3H), 0.90 (d, J=6.3, 3H), 1.00-1.08 (m,1H), 1.29-1.34 (m, 3H), 1.92-1.97 (m, 1H), 2.35 (s, 3H), 2.50 (t, J=6.7,2H), 2.80-3.09 (m, 4H), 3.52 (d, J=5.1, 2H), 3.60-3.62 (t, J=6.8, 1H),7.22 (d, J=7.8, 2H), 7.63 (d, J=7.8, 2H).

Step B. Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-(4-methylbenzenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (400 mg,1.2 mmol, step A) following the indications of general procedure Bdusing Nα-(4-methylbenzenesulfonyl)-L-tryptophan (450 mg, 1.3 mmol,example 2, step A), EDC (290 mg, 1.5 mmol) and HOBt (100 mg, 1.5 mmol).The crude material was purified by flash chromatography using a solventgradient from 39:1 to 19:1 CH₂Cl₂/MeOH. The product was isolated as ayellowish oil (420 mg, 48% yield).

¹H NMR (CDCl₃): δ0.93 (t, J=6.8, 6H), 0.98 (m, 1H), 1.26 (m, 3H), 1.42(m, 1H), 1.93 (m, 4H), 2.38 (s, 3H), 2.42 (s, 3H), 2.91 (m, 1H), 3.05(m, 3H), 3.13 (m, 2H), 3.56 (m, 2H), 3.66 (m, 1H), 3.91 (d, J=5.6, 1H),5.08 (d, J=5.9,1H), 6.32 (t, J=4.1, 1H), 6.95 (s, 1H), 7.02 (t, J=7.5,1H), 7.12 (d, J=7.4, 2H), 7.19 (t, J=7.5, 1H), 7.29 (d, J=8.6, 2H), 7.35(d, J=7.8, 1H), 7.51 (d, J=7.3, 2H), 7.72 (d, J=7.9, 2H), 8.54 (s, 1H).LC-MS: 681 (M−H)⁻, 98% pure.

Step C. Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-(4-methylbenzenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanal

The title compound was prepared from the previous derivative (400 mg,0.59 mmol) by oxidation under Swem's reaction conditions using oxalylchloride (110 μL, 1.2 mmol), DMSO (166 μL, 2.4 mmol), and triethylamine(2 mL) (Organic Syntheses, Collective Volume VII, p. 258-263). The crudematerial was purified by flash chromatography using a solvent gradientfrom 39:1 to 19:1 CH₂Cl₂/MeOH. The product was isolated as a yellowishsolid (330 mg, 83% yield).

¹H NMR (DMSO-d₆): δ0.82 (t, J=7.6, 6H), 0.92 (m, 1H), 1.09 (m, 1H), 1.31(m, 3H), 1.72 (m, 1H), 1.83 (m, 1H), 2.33 (s, 3H), 2.37 (s, 3H), 2.73(d, J=4.8, 1H), 2.96 (m, 5H), 4.08 (t, J=6.1, 1H), 4.72 (m, 1H), 6.93(t, J=7.4, 1H), 6.99 (t, J=7.3, 1H), 7.15 (d, J=7.6, 1H), 7.37 (m, 6H),7.61 (d, J=7.5, 1H), 7.72 (d, J=7.6, 2H), 8.02 (t, J=5.7, 1H), 9.51 (s,1H), 11.35 (s, 1H).

Example 117 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-pivaloyl-S-tryptophanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-pivaloyl-L-tryptophan (example 100, step A). Purification by HPLCgave 85 mg (48%) of the desired material.

¹H NMR (CDCl₃): δ0.90-0.98 (m, 6H), 1.09-1.16 (m, 1H), 1.23 (s, 9H),1.25-1.29 (m, 1H), 1.45-1.52 (m, 1H), 1.92-1.97 (m, 1H), 2.38 (s, 3H),2.82-3.10 (m, 4H), 3.11-3.26 (m, 3H), 3.51-3.55 (m, 1H), 3.51 (d, J=7.0,2H), 4.64 (br s, 1H), 6.97-7.21 (m, 6H), 7.35 (q, J=6.7, 1H), 7.61 (d,J=7.1, 1H), 7.69 (d, J=7.6, 1H). LC-MS: 611.2 (M−H)⁻, 95% pure.

Example 118 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-acetyl-S-phenylalanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-acetyl-L-phenylalanine (example 30, step A). Purification by HPLCgave 82 mg (51%) of the desired material.

¹H NMR (CDCl₃): δ0.79 (d, J=6.3, 3H), 0.82 (d, J=6.3, 3H), 1.08-1.11 (m,2H), 1.23-1.31 (m, 2H), 1.45-1.52 (m, 1H), 1.92-1.97 (m, 1H), 1.98 (d,J=9.0, 3H), 2.38 (s, 3H), 2.84-3.09 (m, 6H), 3.51 (d, J=7.1, 2H),3.61-3.64 (m, 1H), 4.51 (q, J=6.9, 1H), 7.09-7.26 (m, 7H), 7.73 (d,J=8.1, 2H). LC-MS: 530.4 (M−H)⁻, 99% pure.

Example 119 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-pivaloyl-S-phenylalanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-pivaloyl-L-phenylalanine (example 52, step A). Purification by HPLCgave 65 mg (37%) of the desired material.

¹H NMR (CDCl₃): δ0.79 (d, J=6.3, 3H), 0.82 (d, J=6.3, 3H), 1.08-1.11 (m,2H), 1.13 (s, 9H), 1.23-1.31 (m, 2H), 1.45-1.52 (m, 1H), 1.92-1.97 (m,1H), 2.38 (s, 3H), 2.84-3.09 (m, 6H), 3.51 (d, J=7.1, 2H), 3.2 (t,J=6.1, 1H), 4.51 (q, J=6.9, 1H), 7.09-7.26 (m, 7H), 7.73 (d, J=7.9, 2H).LC-MS: 572.2 (M−H)⁻, 99% pure.

Example 120 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-(4-morpholinecarbonyl)-S-phenylalanyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-(4-morpholinecarbonyl)-L-phenylalanine (example 51, step A).Purification by HPLC gave 69 mg (38%) of the desired material.

¹H NMR (CDCl₃): δ0.79 (d, J=6.3, 3H), 0.82 (d, J=6.3, 3H), 1.08-1.11 (m,2H), 1.23-1.31 (m, 2H), 1.45-1.52 (m, 1H), 1.92-1.97 (m, 1H), 2.39 (s,3H), 2.84-3.09 (m, 6), 3.21-3.31 (m, 4H), 3.45-3.64 (m, 7H), 4.48 (br s,1H), 7.09-7.26 (m, 7H), 7.73 (d, J=8.1, 2H). LC-MS: 601.1 (M−H)⁻, 99%pure.

Example 121 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-(4-acetylaminobenzenesulfonyl)-S-phenylalanyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-(4-acetamidobenzenesulfonyl)-L-phenylalanine (example 3, step A).Purification by HPLC gave 103 mg (49%) of the desired material.

¹H NMR (CDCl₃): δ0.79 (d, J=6.3, 3H), 0.82 (d, J=6.3, 3H), 1.08-1.11 (m,2H), 1.23-1.31 (m, 2H), 1.45-1.52 (m, 1H), 1.92-1.97 (m, 1H), 2.20 (s,3H), 2.38 (s, 3H), 2.85-3.02 (m, 6H), 3.51 (d, J=7.1, 2H), 3.64 (q,J=6.4, 1H), 3.88 (q, J=6.9, 1H), 6.89 (br s, 2H), 7.19 (br s, 3H), 7.24(d, J=8.0, 2H), 7.54-7.61 (m, 4H), 7.73 (d, J=8.1, 2H). LC-MS: 685.2(M−H)⁻, 99% pure.

Example 122 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[Nα′-(2-thiophenesulfonyl)-S-phenylalanyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-(2-thiophenesulfonyl)-L-phenylalanine (example 24, step A).Purification by HPLC gave 69 mg (36%) of the desired material.

¹H NMR (CDCl₃): δ0.79 (d, J=6.3, 3H), 0.82 (d, J=6.3, 3H), 1.08-1.11 (m,2H), 1.28-1.35 (m, 2H), 1.45-1.52 (m, 1H), 1.92-1.97 (m, 1H), 2.39 (s,3H), 2.85-3.18 (m, 6H), 3.51 (d, J=7.0, 2H), 3.64 (q, J=6.4, 1H), 3.94(q, J=6.9, 1H), 6.91-6.98 (m, 3H), 7.19 (br s, 3H), 7.24 (d, J=8.0, 2H),7.35 (s, 1H), 7.52 (d, J=4.1, 1H), 7.73 (d, J=8.1, 2H). LC-MS: 634.2(M−H)⁻, 99% pure.

Example 123 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-benzenesulfonyl-S-phenylalanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-benzenesulfonyl-L-phenylalanine (example 8, step A). Purification byHPLC gave 68 mg (36%) of the desired material.

¹H NMR (CDCl₃): δ0.79 (d, J=6.3, 3H), 0.82 (d, J=6.3, 3H), 1.08-1.11 (m,2H), 1.28-1.35 (m, 2H), 1.45-1.52 (m, 1H), 1.92-1.97 (m, 1H), 2.41 (s,3H), 2.85-3.18 (m, 6H), 3.51 (d, J=7.0, 2H), 3.64 (q, J=6.4, 1H), 3.84(q, J=6.9, 1H), 6.88 (d, J=6.6, 2H), 7.14-7.19 (m, 3H), 7.24 (d, J=8.0,2H), 7.35 (t, J=6.8, 2H), 7.52 (t, J=7.1, 1H), 7.60 (d, J=6.8, 2H), 7.72(d, J=8.0, 2H). LC-MS: 628.2 (M−H)⁻, 99% pure.

Example 124 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-(4-methylbenzenesulfonyl)-S-4-nitrophenylalanyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-(4-methylbenzenesulfonyl)-L-4-nitrophenylalanine (example 26, stepA). Purification by HPLC gave 101 mg (48%) of the desired material.

¹H NMR (CDCl₃): δ0.89-0.97 (m, 6H), 1.18-1.21 (m, 2H), 1.35-1.45 (m,3H), 1.92-1.97 (m, 1H), 2.33 (s, 3H), 2.39 (s, 3H), 2.85-3.01 (m, 3H),3.15-3.24 (m, 3H), 3.51 (d, J=7.0, 2H), 3.71 (q, J=6.4, 1H), 3.98 (br s,1H), 7.01 (d, J=7.6, 2H), 7.19 (d, J=8.0, 2H), 7.24 (d, J=8.0, 2H), 7.39(d, J=7.7, 2H), 7.73 (d, J=8.1, 2H), 7.88 (d, J=7.9, 2H). LC-MS: 687.8(M−H)⁻, 99% pure.

Example 125 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-(4-fluorobenzenesulfonyl)-S-phenylalanyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-(4-fluorobenzenesulfonyl)-L-phenylalanine (example 18, step A).Purification by HPLC gave 66 mg (34%) of the desired material.

¹H NMR (CDCl₃): δ0.79 (d, J=6.3, 3H), 0.82 (d, J=6.3, 3H), 1.09-1.16 (m,2H), 1.35-1.39 (m, 2H), 1.45-1.52 (m, 1H), 1.92-1.97 (m, 1H), 2.42 (s,3H), 2.82-3.18 (m, 6H), 3.51 (d, J=7.0, 2H), 3.64 (q, J=6.3, 1H), 3.79(q, J=6.9, 1H), 6.91 (d, J=7.0, 2H), 7.00 (t, J=6.8, 2H), 7.15-7.21 (m,3H), 7.35 (s, 1H), 7.52 (d, J=7.1, 1H), 7.54 (q, J=6.6, 2H), 7.73 (d,J=8.1, 2H). LC-MS: 647.6 (M−H)⁻, 99% pure.

Example 126 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-(4-methoxybenzenesulfonyl)-S-phenylalanyl]-2,6-diaminohexanol

Step A. Preparation of Nα-(4-methoxybenzenesulfonyl)-L-phenylalanine

L-phenylalanine was reacted with 4-methoxybenzenesulfonyl chloride underthe conditions used in general procedure A giving the title compoundwhich was used without purification in the next step.

Step B. Preparation of (2S,2′S)2-Nisobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-(4-methoxybenzenesulfonyl)-S-phenylalanyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diamninohexanol (example116, step A) following the indications of general procedure Bd usingNα-(4-methoxybenzenesulfonyl)-L-phenylalanine (this example, step A).Purification by HPLC gave 107 mg (53%) of the desired material.

¹H NMR (CDCl₃): δ0.79 (d, J=6.3, 3H), 0.82 (d, J=6.3, 3H), 1.09-1.16 (m,2H), 1.35-1.39 (m, 2H), 1.45-1.52 (m, 1H), 1.94-1.97 (m, 1H), 2.40 (s,3H), 2.82-3.18 (m, 6H), 3.51 (d, J=7.0, 2H), 3.64 (q, J=6.3, 1H), 3.79(q, J=6.9, 1H), 3.92 (s, 3H), 6.87 (d, J=6.9, 2H), 6.89 (d, J=7.0, 2H),7.15-7.21 (m, 3H), 7.35 (d, J=7.1, 2H), 7.50 (d, J=7.1, 1H), 7.73 (d,J=8.1, 2H). LC-MS: 658.3 (M−H)⁻, 99% pure.

Example 127 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-(4-trifluoromethylbenzenesulfonyl)-S-phenylalanyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-(4-trifluoromethylbenzenesulfonyl)-L-phenylalanine (example 23, stepA). Purification by HPLC gave 49 mg (23%) of the desired material.

¹H NMR (CDCl₃): δ0.79 (d, J=6.3, 3H), 0.82 (d, J=6.3, 3H), 1.09-1.16 (m,2H), 1.25-1.40 (m, 4H), 1.92-1.97 (m, 1H), 2.44 (s, 3H), 2.78-3.18 (m,6H), 3.51 (d, J=6.8, 2H), 3.64 (q, J=6.3, 1H), 3.84 (q, J=5.9, 1H), 6.91(d, J=7.0, 2H), 7.03-7.21 (m, 3H), 7.30 (d, J=6.8, 2H), 7.52 (d, J=7.1,2H), 7.60 (d, J=6.9, 2H), 7.71 (d, J=7.1, 2H). LC-MS: 696.2 (M−H)⁻, 99%pure.

Example 128 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-benzenesulfonyl-S-tryptophanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-benzenesulfonyl-L-tryptophan (example 4, step A). Purification byHPLC gave 93 mg (46%) of the desired material.

¹H NMR (CDCl₃): δ0.84 (d, J=6.3, 3H), 0.89 (d, J=6.3, 3H), 0.94-1.03 (m,1H), 1.09-1.16 (m, 2H), 1.46-1.49 (m, 1H), 1.93-1.98 (m, 1H), 2.39 (s,3H), 2.82-3.15 (m, 6H), 3.51 (d, J=6.8, 2H), 3.64 (q, J=6.3, 1H), 3.94(q, J=5.5, 1H), 6.95 (t, J=4.5, 1H), 7.19 (t, J=4.5, 1H), 7.23-7.31 (m,6H), 7.42 (t, J=4.5, 1H), 7.60 (d, J=6.8, 2H), 7.73 (d, J=6.8, 2H).LC-MS: 667.5 (M−H)⁻, 99% pure.

Example 129 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-(2-thiophenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-(2-thiophenesulfonyl)-L-tryptophan (example 53, step A). Purificationby HPLC gave 81 mg (40%) of the desired material.

¹H NMR (CDCl₃): δ0.84 (d, J=6.3, 3H), 0.89 (d, J=6.3, 3H), 0.94-1.03 (m,1H), 1.09-1.16 (m, 2H), 1.46-1.49 (m, 1H), 1.93-1.98 (m, 1H), 2.40 (s,3H), 2.82-3.15 (m, 6H), 3.51 (d, J=6.8, 2H), 3.64 (q, J=6.3, 1H), 4.05(t, J=7.2, 1H), 6.89 (t, J=4.6, 1H), 6.97 (s, 1H), 7.03 (t, J=4.5, 1H),7.19 (t, J=4.5, 1H), 7.23-7.28 (m, 3H), 7.34-7.42 (m, 4H), 7.73 (d,J=6.8, 2H). LC-MS: 674.2 (M−H)⁻, 99% pure.

Example 130 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-(4-nitrobenzenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-(4-nitrobenzenesulfonyl)-L-tryptophan (example 5, step A).Purification by HPLC gave 101 mg (47%) of the desired material.

¹H NMR (CDCl₃): δ0.90-0.98 (m, 6H), 1.09-1.16 (m, 2H), 1.35-1.39 (m,2H), 1.45-1.52 (m, 1H), 1.92-1.97 (m, 1H), 2.46 (s, 3H), 2.82-3.10 (m,4H), 3.11-3.26 (m, 3H), 3.51 (d, J=7.0, 2H), 3.74 (q, J=6.3, 1H), 3.95(br s, 1H), 6.91 (t, J=7.0, 1H), 6.97 (s, 1H), 7.01 (t, J=6.8, 1H), 7.15(d, J=6.6, 1H), 7.20-7.30 (m, 3H), 7.52 (d, J=7.1, 1H), 7.71 (d, J=6.9,2H), 7.79 (d, J=7.1, 2H). LC-MS: 713.1 (M−H)⁻, 99% pure.

Example 131 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-acetyl-S-tryptophanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-acetyl-L-tryptophan (example 99, step A). Purification by HPLC gave68 mg (39%) of the desired material.

¹H NMR (CDCl₃): δ0.90-0.98 (m, 6H), 1.09-1.16 (m, 1H), 1.25-1.29 (m,1H), 1.45-1.52 (m, 1H), 1.92-1.97 (m, 1H), 2.0 (s, 3H), 2.38 (s, 3H),2.82-3.10 (m, 4H), 3.11-3.26 (m, 3H), 3.51 (d, J=7.0, 2H), 3.64 (q,J=6.3, 1H), 3.75 (br s, 1H), 6.97-7.21 (m, 6H), 7.35 (q, J=6.7, 1H),7.61 (d, J=7.1, 1H), 7.69 (d, J=7.6, 1H). LC-MS: 569.1 (M−H)⁻, 98% pure.

Example 132 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methoxyphenylacetyl)-L-phenylalanyl]-L-lysine

Step A. Preparation of Nα-(4-methoxyphenylacetyl)-L-phenylalanine

L-phenylalanine was reacted with 4-methoxyphenylacetyl chloride underthe conditions used in general procedure A giving the title compoundwhich was recrystallised neat (80%) and used as such in the next step.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methoxyphenylacetyl)-L-phenylalanyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine hydrochloride (100 mg,0.25 mmol, example 1, step E) as described in general procedure Bc usingNα-(4-methoxyphenylacetyl)-L-phenylalanine (80 mg, 0.25 mmol) which wasprepared in step A of this example. The final product was trituratedwith ether to yield 85 mg (18%) of the desired material.

¹H NMR (CDCl₃): δ0.83 (d, J=6.9, 6H), 1.08-1.11 (m, 2H), 1.33-1.55 (m,2H), 1.45-1.52 (m, 1H), 1.79-1.89 (m, 2H), 2.36 (s, 3H), 2.85-3.27 (m,6H), 3.55 (s, 2H), 3.79 (s, 3H), 4.21 (s, 2H), 4.53 (t, J=5.9, 1H), 6.79(d, J=8.2, 2H), 6.99-7.09 (m, 3H), 7.15-7.26 (m, 7H), 7.73 (d, J=8.1,2H). LC-MS: 650.1 (M−H)⁻, 98% pure.

Example 133 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-dihydrocinnamoyl-L-tryptophanyl)-L-lysine

Step A. Preparation of Nα-dihydrocinnamoyl-L-tryptophan

L-tryptophan was reacted with Nα-dihydrocinnamoyl chloride under theconditions used in general procedure A giving the title compound whichwas recrystallised from DCM (55%). This material was used withoutfurther purification in the next step.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-dihydrocinnamoyl-L-tryptophanyl)-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine hydrochloride (400 mg,1.0 mmol, example 1, step E) as described in general procedure Bc usingNα-dihydrocinnamoyl-L-tryptophan (360 mg, 1.0 mmol) which was preparedin step A of this example. The final product was triturated with etherto yield 612 mg (85% pure) of the crude material. Purification of 100 mgof the crude material by HPLC gave 45 mg (45%) of pure adduct.

¹H NMR (CDCl₃): δ0.82 (d, J=6.8, 3H), 0.84 (d, J=6.8, 3H), 1.08-1.11 (m,2H), 1.18-1.22 (m, 2H), 1.45-1.52 (m, 1H), 1.94-1.99 (m, 2H), 2.34 (s,3H), 2.48-2.52 (m, 2H), 2.89-3.09 (m, 4H), 3.15-3.18 (m, 2H), 4.29 (brs, 1H), 4.77 (br s, 1H), 6.95 (t, J=7.2, 1H), 7.04-7.34 (m, 8H), 7.36(d, J=8.1, 1H), 7.69 (d, J=7.2, 1H), 7.99 (d, J=8.1, 2H). LC-MS: 673.1(M−H)⁻, 98% pure.

Example 134 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-tert-butoxycarbonyl-S-2-pyridylalanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-tert-butoxycarbonyl-L-2-pyridylalanine (Peptech Corporation).Purification by HPLC gave 54 mg (30%) of the desired material.

¹H NMR (CDCl₃): δ0.90-0.98 (m, 6H), 1.09-1.16 (m, 1H), 1.25-1.29 (m,2H), 1.45 (s, 9H), 1.92-1.97 (m, 1H), 2.38 (s, 3H), 2.82-3.10 (m, 4H),3.26 (br s, 2H), 3.51 (d, J=7.0, 2H), 3.64 (q, J=6.3, 1H), 4.55 (br s,1H), 7.18-7.38 (m, 4H), 7.70 (t, J=6.8, 1H), 7.74 (d, J=7.5, 2H), 8.45(s, 1H). LC-MS: 591.3 (M+H)⁺, 98% pure.

Example 135 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-tert-butoxycarbonyl-S-3-pyridylalanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-tert-butoxycarbonyl-L-3 -pyridylalanine (Peptech Corporation).Purification by HPLC gave 45 mg (25%) of the desired material.

¹H NMR (CDCl₃): δ0.90-0.98 (m, 6H), 1.09-1.16 (m, 1H), 1.25-1.29 (m,2H), 1.45 (s, 9H), 1.92-1.97 (m, 1H), 2.37 (s, 3H), 2.82-3.30 (m, 6H),3.51 (d, J=7.0, 2H), 3.64 (q. K=6.3, 1H), 4.35 (br s, 1H), 7.18-7.38 (m,4H), 7.60 (br s, 1H), 7.74 (d, J=7.5, 2H), 8.56 (d, J=6.6, 2H). LC-MS:591.3 (M+H)⁺, 95% pure.

Example 136 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-tert-butoxycarbonyl-S-4-pyridylalanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-tert-butoxycarbonyl-L-4-pyridylalanine (Peptech Corporation).Purification by HPLC gave 65 mg (36%) of the desired material.

¹H NMR (CDCl₃): δ0.90-0.98 (m, 6H), 1.09-1.16 (m, 1H), 1.25-1.29 (m,2H), 1.49 (s, 9H), 1.92-1.97 (m, 1H), 2.37 (s, 3H), 2.82-3.30 (m, 6H),3.49-3.51 (m, 2H), 3.64 (br s, 1H), 4.35 (br s, 1H), 7.16 (s, 2H),7.18-7.22 (m, 2H), 7.74 (d, J=7.5, 2H), 8.56 (br s, 2H). LC-MS: 591.3(M+H)⁺, 98% pure.

Example 137 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-tert-butoxycarbonyl-S-4-thiazolylalanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-tert-butoxycarbonyl-L-4-thiazolylalanine (Peptech Corporation).Purification by HPLC gave 50 mg (29%) of the desired material.

¹H NMR (CDCl₃): δ0.90-0.98 (m, 6H), 1.09-1.16 (m, 1H), 1.25-1.29 (m,2H), 1.49 (s, 9H), 1.92-1.97 (m, 1H), 2.37 (s, 3H), 2.82-3.30 (m, 6H),3.49-3.51 (m, 2H), 3.64 (br s, 1H), 4.35 (br s, 1H), 7.06 (s, 1H),7.18-7.22 (m, 3H), 7.74 (d, J=7.5, 2H). LC-MS: 697.3 (M+H)⁺, 99% pure.

Example 138 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-tert-butoxycarbonyl-S-2-fluorophenylalanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-tert-butoxycarbonyl-L-2-fluorophenylalanine (Peptech Corporation).Purification by HPLC gave 62 mg (34%) of the desired material.

¹H NMR (CDCl₃): δ0.79 (d, J=6.3, 3H), 0.82 (d, J=6.3, 3H), 1.08-1.11 (m,2H), 1.29-1.34 (m, 3H), 1.41 (s, 9H), 1.92-1.97 (m, 1H), 2.38 (s, 3H),2.84-3.09 (m, 6H), 3.51 (d, J=7.1, 2H), 3.72 (t, J=6.1, 1H), 4.35 (br s,1H), 7.00-7.08 (m, 2H), 7.14-7.28 (m, 4H), 7.73 (d, J=7.9, 2H). LC-MS:608.4 (M+H)⁺, 99% pure.

Example 139 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-tert-butoxycarbonyl-S-3-fluorophenylalanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-tert-butoxycarbonyl-L-3-fluorophenylalanine (Peptech Corporation).Purification by HPLC gave 88 mg (48%/) of the desired material.

¹H NMR (CDCl₃): δ0.85 (d, J=6.3, 3H) 0.90 (d, J=6.3, 3H), 1.08-1.11 (m,2H), 1.29-1.34 (m, 3H), 1.41 (s, 9H), 1.92-1.97 (m, 1H), 2.41 (s, 3H),2.84-3.09 (m, 6H), 3.51 (d, J=7.1, 2H), 3.62 (t, J=6.1, 1H), 4.35 (br s,1H), 6.91 (d, J=6.6, 2H), 7.00 (d, J=6.7, 2H), 7.14-7.28 (m, 3H), 7.73(d, J=7.9, 2H). LC-MS: 608.4 (M+H)⁺, 99% pure.

Example 140 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-tert-butoxycarbonyl-S-4-fluorophenylalanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-tert-butoxycarbonyl-L-4-fluorophenylalanine (Peptech Corporation).Purification by HPLC gave 65 mg (35%) of the desired material.

¹H NMR (CDCl₃): δ0.85 (d, J=6.3, 3H), 0.90 (d, J=6.3, 3H), 1.08-1.11 (m,2H), 1.29-1.34 (m, 3H), 1.41 (s, 9H), 1.92-1.97 (m, 1H), 2.41 (s, 3H),2.84-3.09 (m, 6H), 3.51 (d, J=7.1, 2H), 3.60-3.62 (m, 1H), 4.24 (br s,1H), 6.94 (t, J=7.6, 2H), 7.14-7.18 (m, 2H), 7.20-7.30 (m, 2H), 7.73 (d,J=7.9, 2H). LC-MS: 608.4 (M+H)⁺, 99% pure.

Example 141 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-tert-butoxycarbonyl-S-1,2,3,4-tetrahydronorharman-3-carbonyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-tert-butoxycarbonyl-L-1,2,3,4-tetrahydronorharman-3-carboxylic acid(Peptech Corporation). Purification by HPLC gave 75 mg (39%) of thedesired material.

¹H NMR (CDCl₃): δ0.85 (d, J=6.3, 3H), 0.90 (d, J=6.3, 3H), 1.08-1.11 (m,2H), 1.29-1.34 (m, 3H), 1.36 (s, 9H), 1.92-1.97 (m, 1H), 2.35 (s, 3H),2.40 (t, J=6.7, 1H), 2.80-3.09 (m, 6H), 3.52 (d, J=5.1, 2H), 3.60-3.62(m, 1H), 4.23 (br s, 1H), 7.28 (d, J=7.8, 2H), 7.73 (d, J=7.9, 2H).LC-MS: 641.5 (M+H)⁺, 99% pure.

Example 142 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-tert-butoxycarbonyl-S-4-tert-butylphenylalanyl)-2,6-diaminohexanol

The title product was prepared from (25)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-tert-butoxycarbonyl-L-4-tert-butylphenylalanine (PeptechCorporation). Purification by HPLC gave 81 mg (42%) of the desiredmaterial.

¹H NMR (CDCl₃): δ0.85 (d, J=6.3, 3H), 0.90 (d, J=6.3, 3H), 1.08-1.11 (m,2H), 1.29-1.34 (m, 3H), 1.36 (s, 9H), 1.92-1.97 (m, 1H), 2.40 (s, 3H),2.80-3.29 (m, 8H), 3.52 (d, J=5.1, 2H), 3.60-3.62 (m, 1H), 4.23 (br s,1H), 6.90-7.28 (m, 5H), 7.55 (d, J=6.6, 1H)), 7.73 (d, J=7.9, 2H).LC-MS: 646.5 (M+H)⁺, 99% pure.

Example 143 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-tert-butoxycarbonyl-S-pentafluorophenylalanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-tert-butoxycarbonyl-L-pentafluorophenylalanine (Peptech Corporation).Purification by HPLC gave 55 mg (27%) of the desired material.

¹H NMR (CDCl₃): δ0.85 (d, J=6.3, 3H), 0.90 (d, J=6.3, 3H), 1.08-1.11 (m,2H), 1.29-1.34 (m, 3H), 1.41 (s, 9H), 1.92-1.97 (m, 1H), 2.41 (s, 3H),2.80-3.09 (m, 6H), 3.51 (d, J=7.1, 2H), 3.60-3.62 (m, 1H), 4.31 (br s,1H), 7.09 (s, 2H), 7.10-7.28 (m, 5H), 7.68 (d, J=7.5, 2H). LC-MS: 680.2(M+H)⁺, 99% pure.

Example 144 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-tert-butoxycarbonyl-S-4-(9-fluorenemethoxycarbonylaminomethyl)phenylalanyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-tert-butoxycarbonyl-L-4-(9-fluorenemethoxycarbonylaminomethyl)phenylalanine(Peptech Corporation). Purification by HPLC gave 41 mg (16%) of thedesired material.

LC-MS: 842.8 (M+H)⁺, 99% pure.

Example 145 Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-[N′α-(4-methylbenzenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanol

Step A. Preparation ofNα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-Nε-benzyloxycarbonyl-L-lysinemethyl ester

This product was prepared following the procedure described for thepreparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-benzyloxycarbonyl-L-lysinemethyl ester (example 65, step B) using 4-nitrobenzenesulfonyl chlorideinstead of4-methylbenzenesulfonyl chloride. The yield of this reactionwas 42%.

¹H NMR (CDCl₃): δ0.83 (d, J=6.6, 3H), 0.86 (d, J=6.5, 3H), 1.35-1.69 (m,5H), 1.88-2.00 (m, 2H), 2.90 and 3.04 (ABX, J=14.5, 7.5, 2H), 3.18 (m,2H), 3.49 (s, 3H), 4.45 (t, J=6.0, 1H), 4.83 (s, 1H), 5.10 (s, 2H),7.30-7.40 (m, 5H), 8.00 (d, J=8.5, 2H), 8.33 (d, J=8.5, 2H).

Step B. Preparation of Nα-(4-aminobenzenesulfonyl)-Nα-isobutyl-L-lysinemethyl ester

The title compound was obtained by catalytic hydrogenation ofNα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-Nε-benzyloxycarbonyl-L-lysinemethyl ester (step A) as described for the preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine methyl ester (example65, step C). This compound was used without purification in the nextstep.

Step C. Preparation ofNα-(4-aminobenzenesulfonyl)-Nα-isobutyl-Nε-[N′α-(4-methylbenzenesulfonyl)-L-tryptophanyl]-L-lysinemethyl ester

The title compound was prepared in the same manner than example 69 usingNα-(4-aminobenzenesulfonyl)-Nα-isobutyl-L-lysine methyl ester (step B)as the starting material. The yield of this reaction was 65%.

¹H NMR (DMSO-d₆): δ0.77 (t, J=7.5, 6H), 0.90-1.10 (m, 4H), 1.40 (m, 1H),1.70 (m, 1H), 1.85 (m, 1H), 2.29 (s, 3H), 2.87-3.00 (m, 3H), 3.44 (s,3H), 3.85 (m, 1H), 4.10 (s, 2H), 4.24 (t, J=7.3, 1H), 5.74 (s, 2), 6.60(d, J=8.4, 2H), 6.90 (m, 1H), 7.00 (m, 2H), 7.15 (d, J=7.5, 2H),7.30-7.45 (m, 6H), 7.70 (t, J=5.0, 1H), 7.82 (d, J=8.7, 1H), 10.70 (s,1H).

Step D. Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-[N′α-(4-methylbenzenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanol

Nα-(4-aminobenzenesulfonyl)-Nα-isobutyl-Nε-[N′α-(4-methylbenzenesulfonyl)-L-tryptophanyl]-L-lysinemethyl ester (step C) was reduced with LiAlH₄ following the indicationsof general procedure G. The final product was obtained in 37% yield.

¹H NMR (DMSO-d₆): δ0.80 (d, J=7.8, 3H), 0.83 (d, J=7.6, 3H), 0.88-1.22(m, 5H), 1.48 (m, 1H), 1.85 (m, 1H), 2.30 (s, 3H), 2.65-2.80 (m, 4H),2.85 (dd, J=14.5, 7.5, 1H), 2.93 (dd, J=14.3, 7.6, 1H), 3.20-3.40 (m,2H), 3.45 (m, 1H), 3.88 (m, 1H), 4.60 (t, J=5.0, 1H), 5.90 (s, 2H), 6.60(d, J=8.4, 2H), 6.90 (m, 1H), 7.00 (m, 2H), 7.15 (d, J=7.5, 2H),7.30-7.45 (m, 6H), 7.70 (t, J=5.0, 1H), 7.82 (d, J=8.7, 1H), 10.70 (s,1H).

Example 146 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methoxybenzenesulfonyl)-6-N-(S-tryptophanyl)-2,6-diaminohexanoltrifluoroacetic acid salt

This product was obtained quantitatively by treating (2S,2′S)2-N-isobutyl-2-N-(4-methoxybenzenesulfonyl)-6-N-(Nα-tert-butoxycarbonyl-S-tryptophanyl)-2,6-diaminohexanol(example 147) with TFA in CH₂Cl₂.

¹H NMR (DMSO-d₆): δ0.80 (d, J=7.0, 3H), 0.83 (d, J=7.0, 3H), 0.92-1.60(m, 6H), 1.88 (m, 1H), 2.78-3.00 (m, 4H), 3.05 and 3.20 (ABX, J=14.0,7.0, 2H), 3.22 and 3.30 (ABX, J=14.2, 7.0, 2H), 3.50 (m, 1H), 3.80 (s,3H), 3.90 (m, 1H), 4.35 (m, 1H), 6.95-7.12 (m, 4H), 7.18 (s, 1H), 7.38(d, J=8.2, 1H), 7.60 (m, 1H), 7.70 (d, J=8.2, 2H), 8.05 (br s, 3H), 8.34(m, 1H), 11.0 (s, 1H).

Example 147 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methoxybenzenesulfonyl)-6-N-(N′α-tert-butoxycarbonyl-S-tryptophanyl)-2,6-diaminohexanol

Step A. Preparation ofNα-isobutyl-Nα-(4-methoxybenzenesulfonyl)-Nε-benzyloxycarbonyl-L-lysinemethyl ester

This product was prepared following the procedure described for thepreparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-benzyloxycarbonyl-L-lysinemethyl ester (example 65, step B) using 4-methoxybenzenesulfonylchloride instead of 4-methylbenzenesulfonyl chloride. The yield of thisreaction was 65%.

¹H NMR (CDCl₃): δ0.84 (d, J=6.6, 3H), 0.85 (d, J=6.5, 3H), 1.30-1.65 (m,5H), 1.85-1.95 (m, 2H), 2.90 and 3.04 (ABX, J=14.5, 7.5, 2H), 3.15 (m,2H), 3.51 (s, 3H), 3.85 (s, 3H), 4.35 (t, J=5.5, 1H), 4.80 (br s, 1H),5.09 (s, 2H), 6.94 (d, J=8.6, 2H), 7.30-7.40 (m, 5H), 7.25 (d, J=8.5,2H).

Step B. Preparation ofNα-isobutyl-Nα-(4-methoxybenzenesulfonyl)-L-lysine methyl ester

The title compound was obtained quantitatively by catalytichydrogenation ofNα-isobutyl-Nα-(4-methoxybenzenesulfonyl)-Nε-benzyloxycarbonyl-L-lysinemethyl ester (step A) as described for the preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine methyl ester (example65, step C). This compound was used without purification in the nextstep.

Step C. Preparation ofNα-isobutyl-Nα-(4-methoxybenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-L-tryptophanyl)-L-lysinemethyl ester

The title compound was prepared in the same manner as in the example 65(step D) using Nα-isobutyl-Nα-(4-methoxybenzenesulfonyl)-L-lysine methylester (step B) and commercially availableNα-tert-butoxycarbonyl-L-tryptophan as the starting materials. The yieldof this reaction was 86%.

¹H NMR (DMSO-d₆): δ0.84 (t, J=7.0, 6H), 0.90-1.30 (m, 5H), 1.31 (s, 9H),1.53 (m, 1H), 1.86 (m, 1H), 2.82-3.10 (m, 5H), 3.46 (s, 3H), 3.83 (s,3H), 4.12 (m, 1H), 4.30 (t, J=5.0, 1H), 6.67 (d, J=8.2, 1H), 6.90-7.12(m, 5H), 7.30 (d, J=8.0, 2H), 7.55 (m, 1H), 7.70 (m, 2H), 7.81 (m, 1H).

Step D. Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methoxybenzenesulfonyl)-6-N-(N′α-tert-butoxycarbonyl-S-tryptophanyl)-2,6-diaminohexanol

Nα-isobutyl-Nα-(4-methoxybenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-L-tryptophanyl)-L-lysinemethyl ester (step C) was reduced with LiAlH₄ following the indicationsof general procedure G. The final product was obtained in 81% yield.

¹H NMR (DMSO-d₆): δ0.73 (t, J=7.0, 6H), 0.90-1.27 (m, 5H), 1.31 (s, 9H),1.52 (m, 1H), 1.85 (m, 1H), 2.80 and 3.02 (ABX, J=14.0, 7.2, 2H), 2.90(m, 2H), 3.50 (m, 1H), 3.82 (s, 3H), 4.15 (m, 1H), 4.65 (t, J=5.0, 1H),6.65 (d, J=7.8, 1H), 6.92-7.12 (m, 5H), 7.30 (d, J=7.8, 1H), 7.58 (d,J=8.0, 1H), 7.70 (d, J=7.7, 2H), 7.73 (t, J=5.0, 1H), 10.77 (s, 1H).

Example 148 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methoxybenzenesulfonyl)-6-N-(N′α-pivaloyl-S-tryptophanyl)-2,6-diaminohexanol

Treatment of (2S,2′S)2-N-isobutyl-2-N-(4-methoxybenzenesulfonyl)-6-N-(S-tryptophanyl)-2,6-diaminohexanoltrifluoroacetic acid salt (example 146) with pivaloyl chloride using theconditions described in example 65 (step B) afforded the desired productin 86% yield.

¹H NMR (DMSO-d₆): δ0.80 (d, J=7.0, 3H), 0.83 (d, J=7.0, 3H), 1.02 (s,9H), 1.03-1.30 (m, 5H), 1.53 (m, 1H), 1.88 (m, 1H), 2.80 (dd, J=14.5,7.5, 1H), 2.83-3.10 (m, 5H), 3.27 and 3.32 (ABX, J=14.5, 7.2, 2H), 3.52(m, 1H), 3.82 (s, 3H), 4.46 (m, 1H), 6.90-7.15 (m, 5H), 7.30 (d, J=8.1,1H), 7.56 (d, J=8.0, 1H), 7.71 (d, J=8.2, 2H), 7.74 (t, J=5.0, 1H),10.75 (s, 1H).

Example 149 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methoxybenzenesulfonyl)-6-N-[N′α-(4-methylbenzenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanol

Treatment of (2S,2′S)2-N-isobutyl-2-N-(4-methoxybenzenesulfonyl)-6-N-(S-tryptophanyl)-2,6-diaminohexanoltrifluoroacetic acid salt (example 146) with 4-methylbenzenesulfonylchloride using the conditions described in example 65 (step B) affordedthe desired product in 82% yield.

¹H NMR (DMSO-d₆): δ0.80 (d, J=6.0, 3H), 0.83 (d, J=6.2, 3H), 0.88-1.50(m, 6H), 1.88 (m, 1H), 2.30 (s, 3H), 2.65 (m, 2H), 2.78 (m, 2H), 2.93(m, 2H), 3.22-3.40 (m, 2), 3.50 (m, 1H), 3.82 (s, 3H), 3.86 (m, 1H),4.67 (m, 1H), 6.90 (t, J=7.5, 1H), 7.00-7.18 (m, 6H), 7.28 (d, J=8.3,1H), 7.34 (d, J=8.0, 1H), 7.45 (d, J=7.5, 2H), 7.71 (m, 3H), 7.83 (d,J=7.4, 1H), 10.71 (s, 1H).

Example 150 Preparation ofNα-cyclopentylmethyl-Nα-(4-methoxybenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-L-tryptophanyl)-L-lysinemethyl ester

Step A. Preparation ofNα-cyclopentylmethyl-Nα-(4-methoxybenzenesulfonyl)-Nε-benzyloxycarbonyl-L-lysinemethyl ester

This product was prepared following the procedure described for thepreparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-benzyloxycarbonyl-L-lysinemethyl ester (example 65, steps A and B) usingcyclopentanecarboxaldehyde instead of isobutyraldehyde (for step A) and4-methoxybenzenesulfonyl chloride instead of 4-methylbenzenesulfonylchloride (for step B). The yield of this two-step sequence was 79%.

¹H NMR (CDCl₃): δ1.03-1.45 (m, 6H), 1.48-1.71 (m, 8H), 1.56 (s, 9H),1.92 (m, 1H), 2.18 (m, 1H), 3.03 and 3.13 (ABX, J=13.5, 7.0, 2H), 3.20(m, 2H), 3.53 (s, 3H), 3.87 (s, 3H), 4.40 (t, J=6.5, 1H), 5.11 (s, 2H),6.96 (d, J=8.7, 2H), 7.25-7.40 (m, 6H), 7.77 (d, J=8.7, 2H).

Step B. Preparation ofNα-cyclopentylmethyl-Nα-(4-methoxybenzenesulfonyl)-L-lysine methyl ester

The title compound was obtained quantitatively by catalytichydrogenation ofNα-cyclopentylmethyl-Nα-(4-methoxybenzenesulfonyl)-Nε-benzyloxycarbonyl-L-lysinemethyl ester as described for the preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine methyl ester (example65, step C). This compound was used without purification in the nextstep.

Step C. Preparation ofNα-cyclopentylmethyl-Nα-(4-methoxybenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-L-tryptophanyl)-L-lysinemethyl ester

The title compound was prepared in the same manner as in example 65(step D) usingNα-cyclopentylmethyl-Nα-(4-methoxybenzenesulfonyl)-L-lysine methyl ester(this example, step B) andNα-tert-butoxycarbonyl-L-tryptophan-N-hydroxysuccinimide as the startingmaterials. The yield of the two last reactions was 82%.

¹H NMR (CDCl₃): δ1.08-1.28 (m, 6H), 1.30 (s, 9H), 1.40-1.62 (m, 7H),1.80 (m, 1H), 2.10 (m, 1H), 2.82-3.10 (m, 5H), 3.46 (s, 3H), 3.82 (s,3H), 4.12 (m, 1H), 4.30 (m, 1H), 6.65 (d, J=8.2, 1H), 6.90-7.10 (m, 5H),7.30 (d, J=8.0, 1H), 7.59 (d, J=8.2, 1H), 7.70 (d, J=8.1, 2H), 7.80 (t,J=5.0, 1H), 10.77 (s, 1H).

Example 151 Preparation ofNα-cyclopentylmethyl-Nα-(4-methoxybenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-L-tryptophanyl)-L-lysine

A saponification ofNα-cyclopentylmethyl-Nα-(4-methoxybenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-L-tryptophanyl)-L-lysinemethyl ester (example 150) using the conditions described in example 65(step E) afforded the desired material quantitatively.

¹H NMR (DMSO-d₆): δ1.08-1.28 (m, 6H), 1.21 (s, 9H), 1.40-1.65 (m, 7H),1.80 (m, 1H), 2.18 (m, 1H), 2.88-3.10 (m, 6H), 3.82 (s, 3H), 4.12 (m,1H), 4.20 (m, 1H), 6.62 (d, J=7.8, 1H), 6.90-7.15 (m, 5H), 7.3 (d,J=8.0, 1H), 7.55 (d, J=8.0, 1H), 7.70 (d, J=8.7, 2H), 7.80 (d, J=5.1,1H), 10.77 (s, 1H), 12.70 (br s, 1H).

Example 152 Preparation of (2S,2′S)2-N-cyclopentylmethyl-2-N-(4-methoxybenzenesulfonyl)-6-N-(S-tryptophanyl)-2,6-diaminohexanoltrifluoroacetic acid salt

This product was obtained quantitatively by treating (2S,2′S)2-N-cyclopentylmethyl-2-N-(4-methoxybenzenesulfonyl)-6-N-(N′α-tert-butoxycarbonyl-S-tryptophanyl)-2,6-diaminohexanol(example 153) with TFA in CH₂Cl₂.

¹H NMR (DMSO-d₆): δ1.10-1.38 (m, 7H), 1.40-1.68 (m, 7H), 2.18 (m, 1H),2.84-3.02 (m, 4H), 3.10 and 3.18 (ABX, J=14.0, 7.0, 2H), 3.30 (m, 2H),3.50 (m, 1H), 3.81 (s, 3H), 3.90 (m, 1H), 4.35 (m, 1H), 6.98-7.10 (m,4H), 7.20 (s, 1H), 7.39 (d, J=8.3, 1H), 7.60 (m, 1H), 7.70 (d, J=8.2,2H), 8.10 (br s, 3H), 8.36 (m, 1H), 11.0 (s, 1H).

Example 153 Preparation of (2S,2′S)2-N-cyclopentylmethyl-2-N-(4-methoxybenzenesulfonyl)-6-N-(N′α-tert-butoxycarbonyl-S-tryptophanyl)-2,6-diaminohexanol

Nα-cyclopentylmethyl-Nα-(4-methoxybenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-L-tryptophanyl)-L-lysinemethyl ester (example 150) was reduced with LiAlH₄ following theindications of general procedure G. The final product was obtained with72% yield.

¹H NMR (DMSO-d₆): δ0.95-1.28 (m, 7H), 1.30 (s, 9H), 1.42-1.65 (m, 6H),2.15 (m, 1H), 2.85-3.10 (m, 6H), 3.35 (m, 1H), 3.53 (m, 1H), 3.80 (s,3H), 4.15 (m, 1H), 4.65 (m, 1H), 6.75 (d, J=8.2, 1H), 6.90-7.12 (m, 5H),7.30 (d, J=8.0, 1H), 7.55 (d, J=8.4, 1H), 7.70 (d, J=8.0, 2H), 7.73 (t,J=5.0, 1H).

Example 154 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methylbenzenesulfonyl)-L-phenylalanyl]-L-ornithine

The title compound was prepared using the conditions described inexample 65 (steps A, B, C, D and E) with, as the starting materials,Nε-benzyloxycarbonyl-L-ornithine methyl ester hydrochloride and, forstep D, Nα-(4-methylbenzenesulfonyl)-L-phenylalanine.

¹H NMR (DMSO-d₆): δ0.77 (d, J=6.5, 3H), 0.80 (d, J=6.8, 3H), 1.15-1.25(m, 2H), 1.40 (m, 1H), 1.70 (m, 1H), 1.90 (m, 1H), 2.33 (s, 3H), 2.36(s, 3H), 2.60-3.00 (m, 5H), 3.85 (m, 2H), 4.19 (m, 2H), 4.19 (m, 1H),7.08-7.26 (m, 6H), 7.38 (d, J=8.2, 2H), 7.45 (m, 2H), 7.70 (d, J=8.3,2H), 7.35 (t, J=5.0, 1H), 7.90 (d, J=8.5, 1H), 8.16 (d, J=8.6, 1H),12.70 (br s, 1H).

Example 155 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α,N′α-dibenzylglycyl)-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzylglycyl)-L-lysine(100 mg, 0.20 mmol, example 109) by following the indications of generalprocedure H using Et₃N (70 μL, 0.50 mmol) and benzyl bromide (0.10 mL,0.84 mmol). The crude material was purified by preparative HPLC. Theproduct was isolated as a solid (21 mg, 18% yield).

¹H NMR (CDCl₃): δ0.78 (d, J=6.7, 3H), 0.83 (d, J=6.8, 3H), 1.34 (m, 2H),1.45 (m, 1H), 1.65 (m, 2H), 1.91 (m, 2H), 2.39 (s, 3H), 2.86 (dd,J=13.6, 7.3, 1Ha), 2.98 (dd, J=13.6, 7.3, 1Hb), 3.12 (s, 1H), 3.15 (m,2H), 3.64 (s, 4H), 4.43 (t, J=7.0, 1H), 4.93 (s, 2H), 7.13 (s, 1H), 7.16(d, J=7.7, 2H), 7.22-7.37 (m, 10H), 7.66 (d, J=7.6, 2H). LC-MS: 592(M−H)⁻, 99% pure.

Example 156 Preparation of (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-benzyl-N′α-phenylglycyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (350 mg,1.0 mmol, example 116, step A) following the indications of generalprocedure Ba using N-benzyl-N-phenylglycine (241 mg, 1.0 mmol, example114, step A) and N,N-carbonyldiimidazole (195 mg, 1.2 mmol). The crudematerial was purified by preparative HPLC. The product was isolated as asolid (135 mg, 24% yield).

¹H NMR (DMSO-d₆): δ0.83 (t, J=6.8, 6H), 1.05 (m, 2H), 1.26 (m, 3H), 1.55(m, 1H), 1.85 (m, 1H), 2.37 (s, 3H), 2.86 (m, 1H), 2.94 (m, 3H), 3.27(m, 1H), 3.45 (m, 1H), 3.94 (s, 2H), 4.62 (s, 2H), 4.66 (t, J=5.1, 1H),6.57 (d, J=8.3, 2H), 6.61 (t, J=7.5, 1H), 7.11 (t, J=7.5, 2H), 7.25 (m,3 H), 7.31 (t, J=7.4, 2H), 7.35 (d, J=7.8, 2H), 7.66 (d, J=8.2, 2H),8.81 (t, J≅5.0, 1H). LC-MS: 566 (M+H)⁺, 99% pure.

Example 157 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-cyclohexylglycyl)-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-iodoacetyl-L-lysine (520 mg,1.0 mmol, example 105, step B) by following the indications of generalprocedure H using cyclohexylamine (1.0 mL, 8.7 mmol). The crude materialwas purified by preparative HPLC. The product was isolated as a solid(300 mg, 64% yield).

¹H NMR (CDCl₃/MeOD 1:1): δ0.85 (d, J=7.2, 3H), 0.88 (d, J=5.8, 3H), 1.23(m, 1H,), 1.30-1.50 (m, 6H), 1.56 (m, 3H), 1.70 (m, 1H), 1.88 (m, 3H),2.04 (m, 3H), 2.41 (s, 3H), 2.98 (m, 1H), 3.07 (m, 2H), 3.26 (t, J=6.4,2H), 3.66 (s, 2H), 4.27 (t, J=7.3, 1H), 7.31 (d, J=8.4, 2H), 7.75 (d,J=8.8, 2H). LC-MS: 494 (M−H)⁻, 95% pure.

Example 158 Preparation of (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-benzoyl-N′α-benzylglycyl)-2,6-diaminohexanol

Step A. Preparation of N-benzylglycine methyl ester

A solution of methyl bromoacetate (2.0 mL, 20 mmol) in CH₂Cl₂ (25 mL)was treated with benzylamine (3.22 g, 30 mmol). The resulting mixturewas stirred at room temperature for 16 h under an inert atmosphere ofargon. Afterwards, the reaction was quenched with 2N HCl (10 mL). Theproduct was extracted with EtOAc (15 mL, 3×), dried over MgSO₄, filteredand evaporated to an oil. The crude material was purified by flashchromatography using 9:1 hexane/EtOAc. The product was isolated as ayellowish liquid (3.24 g, 86% yield).

¹H NMR (CDCl₃): δ3.44 (s, 2H), 3.74 (s, 3H), 3.82 (s, 2H), 7.27 (m, 1H),7.34 (m, 4H).

Step B. Preparation of N-benzoyl-N-benzylglycine

To a solution of N-benzylglycine methyl ester (800 mg, 4.5 mmol, step A)in THF (10 mL) was added benzoique acid (820 mg, 6.7 mmol) and DCC (1.40g, 6.8 mmol). The reaction mixture was stirred at room temperature for 5h. Then, it was treated with a saturated NaHCO₃ solution and extractedwith EtOAc (30 mL, 3×). The organic phase was evaporated to an oil. Theester intermediate was saponified following the indications of example65 (step E). The crude material was purified by flash chromatographyusing EtOAc:hexane:CHCl₃ (5:5:2) to give 875 mg (73%) of the finalproduct.

¹H NMR (DMSO-d₆): δ 3.84 (s, 0.66H), 4.20 (s, 1.33H), 4.64 (s, 1.33H),4.82 (s, 0.66H), 7.21 (d, J=7.4, 2H), 7.40 (m, 8H), 7.55 (d, J=7.2, 2H),10.0 (s, 1H). LC-MS: 270 (M+H)⁺ and 292 (M+Na)⁺, 98% pure.

Step C. Preparation of (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-benzoyl-N′α-benzylglycyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (350 mg,1.0 mmol, example 116, step A) following the indications of generalprocedure Ba using N-benzoyl-N-benzylglycine (269 mg, 1.0 mmol, step B)and N,N-carbonyldiimidazole (180 mg, 1.1 mmol). The crude material waspurified by preparative HPLC. The product was isolated as a solid (98mg, 17% yield).

¹H NMR (DMSO-d₆): δ 0.83 (t, J=7.1, 6H), 1.05 (m, 2H), 1.26 (m, 3H),1.55 (m, 1H), 1.87 (m, 1H), 2.37 (s, 3H), 2.80-3.00 (m, 4H), 3.52 (m,1H), 3.65 (s, 0.66H), 3.89 (s, 0.33H), 4.49 (s, 0.33H), 4.66 (s, 0.66H),7.20 (m, 1H), 7.29 (m, 3H), 7.36 (d, J=7.3, 2H), 7.43 (m, 5H), 7.67 (d,J=7.5, 2H), 7.83 (m, 1H). LC-MS: 594 (M+H)⁺, 99% pure.

Example 159 Preparation of (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-benzyl-N′α-(4-methylbenzenesulfonyl)glycyl]-2,6-diaminohexanol

Step A. Preparation of N-(4-methylbenzenesulfonyl)-N-benzylglycine

A solution fo N-benzylglycine methyl ester (1.00 g, 5.6 mmol, example158, step A) in DCM (10 mL) was treated with triethylamine (1 mL, 7.2mmol) and 4-methylbenzenesulfonyl chloride (1.28 g, 6.7 mmol). Thereaction mixture was stirred for 3 h at room temperature. Afterwards,the reaction was quenched with 2N HCl (10 mL). The product was extractedwith DCM (15 mL, 3×), dried over MgSO₄, filtered and evaporated. Theester intermediate was saponified following the indications of example65 (step E). The crude material was diluted with 0.5N HCl (150 mL). Theresulting precipitate was filtered off and dried under vacuum to give1.46 g (82%) of the desired material (99% pure).

¹H NMR (DMSO-d₆): δ 2.40 (s, 3H), 3.80 (s, 2H), 4.40 (s, 2H), 7.21 (d,J=7.2, 1H), 7.30 (m, 3H), 7.40 (d, J=7.6 2H), 7.74 (d, J=8.2, 2H), 12.7(s, 1H).

Step B. Preparation of (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-benzyl-N′α-(4-methylbenzenesulfonyl)glycyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (350 mg,1.0 mmol, example 116, step A) following the indications of generalprocedure Ba using N-(4-methylbenzenesulfonyl)-N-benzylglycine (320 mg,1.0 mmol, step A) and N,N-carbonyldiimidazole (180 mg, 1.1 mmol). Thecrude material was purified by preparative HPLC. The product wasisolated as a solid (236 mg, 37% yield).

¹H NMR (DMSO-d₆): δ 0.83 (t, J=7.3, 6H), 1.05 (m, 2H), 1.12 (m, 2H),1.23 (m, 1H), 1.55 (m, 1H), 1.85 (m, 1H), 2.36 (s, 3H), 2.40 (s, 3H),2.80-3.00 (m, 4H), 3.34 (m, 2H), 3.52 (m 1H), 3.65 (s, 2H), 4.39 (s,2H), 7.25 (d, J=7.0, 1H), 7.27-7.40 (m, 7H), 7.60 (t, 4.2, 1H), 7.66 (d,J=8.2, 2H), 7.75 (d, J=7.5, 2H). LC-MS: 644 (M+H)⁺, 90% pure.

Example 160 Preparation of (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-phenethyl-N′α-phenylglycyl]-2,6-diaminohexanol

Step A. Preparation of 2-phenylethyl-4-methylbenzenesulfonate

A solution of 2-phenylethanol (1.22 g, 10 mmol) in THF (10 mL) was addedto a suspension of NaH (440 mg, 11.0 mmol, 60% in oil) in dry THF (5 mL)and stirred for 15 min. The resulting alcolate solution was addeddropwise to a solution of 4-methylbenzenesulfonyl chloride (3.80 g, 20.0mmol) in dry THF (10 mL) and stirred for a period of 1 h. Afterwards,the reaction was quenched with 2N HCl (5 mL). The product was extractedwith EtOAc (25 mL, 3×), dried over MgSO₄, filtered and evaporated to anoil. The crude material was purified by flash chromatography using 9:1hexane/EtOAc. The product was isolated as an oil (1.77 g, 64% yield).

¹H NMR (CDCl₃): δ 2.43 (s, 3H), 2.96 (t, J=7.3, 2H), 4.23 (t, J=7.2,2H), 7.12 (d, J=7.3, 2H), 7.26 (m, 5H), 7.70 (d, J=7.6, 2H).

Step B. Preparation of N-phenethyl-N-phenylglycine

The title compound was prepared from N-phenylglycine (800 mg, 5.30 mmol)as described for the preparation of N-benzyl-N-phenylglycine (example114, step A) using 2-phenylethyl-4-methylbenzenesulfonate (1.61 g, 5.81mmol) instead of benzylbromide. The crude material was purified by flashchromatography using a solvent gradient from 19:1 to 9:1 CH₂Cl₂/MeOH.The product was isolated as a solid (550 mg, 41% yield).

¹H NMR (CDCl₃): δ 2.95 (t, J=7.5, 2H), 3.65 (t, J=7.5, 2H), 3.95 (s,2H), 6.73 (d, J=8.1, 2H), 6.80 (t, J=7.0, 1H), 7.30 (m, 7H).

Step C. Preparation of (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(Nα-phenethyl-N′α-phenylglycyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (320 mg,0.94 mmol, example 116, step A) following the indications of generalprocedure Ba using N-phenethyl-N-phenylglycine (200 mg, 1.0 mmol, stepB) and N,N-carbonyldiimidazole (140 mg, 0.86 mmol). The crude materialwas purified by preparative HPLC. The product was isolated as a hardyoil (32 mg, 7% yield).

¹H NMR (DMSO-d₆): δ 0.83 (t, J=7.1, 6H), 1.05 (m, 2H), 1.25 (m, 3H),1.55 (m, 1H), 1.85 (m, 1H), 2.37 (s, 3H), 2.80-3.01 (m, 4H), 3.52 (m,1H), 3.94 (s, 2H), 4.62 (s, 2H), 4.65 (t, J=4.3, 1H), 6.58 (d, J=7.8, 2H), 6.61 (t, J=7.8, 1 H), 7.11 (t, J=7.3, 2H), 7.25 (m, 3 H), 7.31 (t,J=7.3, 2H), 7.35 (d, J=8.2, 2H), 7.66 (d, J=7.9, 2H), 7.81 (t, J=5.4,1H). LC-MS: 579 (M+H)⁺, 90% pure.

Example 161 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-(4-aminobenzenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-(4-nitrobenzenesulfonyl)-L-tryptophan (example 5, step A). Theintermediate was reduced by catalytic hydrogenation following theconditions of general procedure E. Purification by HPLC gave 30 mg (14%)of the desired material.

¹H NMR (CDCl₃): δ 0.84 (d, J=6.3, 3H), 0.89 (d, J=6.3, 3H), 0.94-1.03(m, 1H), 1.09-1.16 (m, 2H), 1.46-1.49 (m, 1H), 1.93-1.98 (m, 1H), 2.39(s, 3H), 2.82-3.15 (m, 6H), 3.51 (d, J=6.8, 2H), 3.64 (q, J=6.3, 1H),3.89 (q, J=5.5, 1H), 6.48 (d, J=7.8, 2H), 6.95 (t, J=4.5, 1H), 7.19 (t,J=4.5, 1H), 7.23-7.31 (m, 6H), 7.42 (t, J=4.5, 1H), 7.73 (d, J=7.8, 2H).

Example 162 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-(4-methoxyphenylacetyl)-S-phenylalanyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-(4-methoxyphenylacetyl)-L-phenylalanine (example 132, step A).Purification by HPLC gave 25 mg (13%) of the desired material.

¹H NMR (CDCl₃): δ 0.79 (d, J=6.3, 3H), 0.82 (d, J=6.3, 3H), 1.09-1.16(m, 2H), 1.35-1.39 (m, 2H), 1.45-1.52 (m, 1H) 1.94-1.97 (m, 1H), 2.40(s, 3H), 2.82-3.18 (m, 6H), 3.45 (s, 2H), 3.51 (d, J=7.0, 2H), 3.64 (q,J=6.3, 1H), 3.79 (q, J=6.9, 1H), 3.82 (s, 3H), 4.50 (q, J=5.6, 1H), 6.87(d, J=6.9, 2H), 6.89 (d, J=7.0, 2H), 7.15-7.21 (m, 3H), 7.35 (d, J=7.1,2H), 7.50 (d, J=7.1, 1H), 7.73 (d, J=8.1, 2H).

Example 163 Preparation ofNα-(4-aminobenzenesulfonyl)-Nα-isobutyl-Nε-[N′α-(4-methoxyphenylacetyl)-L-phenylalanyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-L-lysine hydrochloride (200 mg,0.45 mmol, example 5, step C) as described in general procedure Bc usingNα-(4-methoxyphenylacetyl)-L-phenylalanine (example 132, step A). Thefinal product,Nα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-Nε-[N′α-(4-methoxyphenylacetyl)-L-phenylalanyl]-L-lysinewas subsequently hydrogenolysed following the indications of generalprocedure E. Purification by HPLC gave the desired material (7 mg, 4%).

¹H NMR (CDCl₃): δ 0.79 (d, J=6.3, 3H), 0.82 (d, J=6.3, 3H), 1.09-1.16(m, 2H), 1.35-1.39 (m, 2H), 1.45-1.52 (m, 1H), 1.94-1.97 (m, 1H), 2.40(s, 3H), 2.82-3.18 (m, 6H), 3.45 (s, 2H), 3.64 (q, J=6.3, 1H), 3.82 (s,3H), 4.33 (t, J=5.5, 1H), 4.50 (q, J=5.6, 1H), 6.87 (d, J=6.9, 2H), 6.89(d, J=7.0, 2H), 7.15-7.21 (m, 3H), 7.35 (d, J=7.1, 2H), 7.50 (d, J=7.1,1H), 7.73 (d, J=8.1, 2H).

Example 164 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methoxyphenylacetyl)-L-tryptophanyl]-L-lysine

Step A. Preparation of Nα-(4-methoxyphenylacetyl)-L-tryptophan

L-tryptophan was reacted with 4-methoxyphenylacetyl chloride under theconditions used in general procedure A giving the title compound whichwas recrystallised from DCM (31%) and used as such in the next step.

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methoxyphenylacetyl)-L-tryptophanyl]-L-lysine

The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine hydrochloride (100 mg,0.25 mmol, example 1, step E) as described in general procedure Bc usingNα-(4-methoxyphenylacetyl)-L-tryptophan (90 mg, 0.25 mmol) which wasprepared in step A of this example. The final product was trituratedwith ether to yield 21 mg (11%) of the desired material.

¹H NMR (CDCl₃): δ 0.79 (d, J=6.3, 3H), 0.82 (d, J=6.3, 3H), 1.09-1.16(m, 2H), 1.35-1.39 (m, 2H), 1.94-1.97 (m, 2H), 2.30 (s, 3H), 2.82-3.18(m, 6H), 3.45 (s, 2H), 3.82 (s, 3H), 4.25-4.20 (m, 1H), 4.50 (q, J=5.6,1H), 6.77 (d, J=6.2, 2H), 6.89 (s, 1H), 6.97 (d, J=6.9, 2H), 7.00-7.11(m, 3H), 7.15-7.21 (m, 3H), 7.35 (d, J=7.1, 2H), 7.50 (d, J=7.1, 1H),7.73 (d, J=8.1, 2H).

Example 165 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-(4-methoxyphenylacetyl)-S-tryptophanyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-(4-methoxyphenylacetyl)-L-tryptophan (example 164, step A).Purification by HPLC gave 20 mg (10%) of the desired material.

¹H NMR (CDCl₃): δ 0.79 (d, J=6.3, 3H), 0.82 (d, J=6.3, 3H), 1.09-1.16(m, 2H), 1.35-1.39 (m, 2H), 1.94-1.97 (m, 2H), 2.30 (s, 3H), 2.82-3.18(m, 6H), 3.45 (s, 2H), 3.51 (d, J=7.0, 2H), 3.82 (s, 3H), 4.50 (q,J=5.6, 1H), 6.77 (d, J=6.2, 2H), 6.89 (s, 1H), 6.97 (d, J=6.9, 2H),7.00-7.11 (m, 3H), 7.15-7.21 (m, 3H), 7.35 (d, J=7.1, 2H), 7.50 (d,J=7.1, 1H), 7.73 (d, J=8.1, 2H).

Example 166 Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-[N′α-(2-thiophenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanol

Step A. Preparation ofNα-isobutyl-Nα-(4-acetamidobenzenesulfonyl)-L-α-amino-ε-caprolactam

Nα-isobutyl-L-α-amino-ε-caprolactam (4.14 g, 21.1 mmol, free base,example 1, step C) was dissolved in DCE (50 mL) and treated withdiisopropylethylamine (6 mL, 0.3 mol) followed by freshly recrystallized4-acetamidobenzenesulfonyl chloride (5.06 g, 21.6 mmol). The mixture wasstirred overnight (TLC shows the reaction to be complete after 2 h). Thesolution was extracted with 1N HCl (50 mL) and the organic layer wasdried and evaporated. The crude material (7.01 g, 83%) was of sufficientpurity to be used as such in the next step.

¹H NMR (CDCl₃): δ 0.93 (d, J=6.0, 3H), 0.96 (d, J=6.0, 3H), 1.39 (t,J=12.0, 1H), 1.85-1.65 (m, 3H), 2.08-2.18 (m and s, 6H), 2.90-2.97 (m1H), 3.00-3.06 (m, 2H), 3.35 (dd, J=14.2, 8.5, 1H), 4.65 (d, J=8.7, 1H),6.3 (s, 1H), 7.42 (d, J=8.8, 2H), 7.6 (d, J=8.8, 2H).

Step B. Preparation of Nα-(4-aminobenzenesulfonyl)-Nα-isobutyl-L-lysinepotassium salt

A mixture ofNα-isobutyl-Nα-(4-acetamidobenzenesulfonyl)-L-α-amino-ε-caprolactam (6.8g, 2 mmol) and 6N HCl (200 mL) was refluxed for 12 h until all solidshad disappeared. Afterwards, the solution was evaporated to dryness. Theresulting solid was dissolved in EtOH (15 mL), neutralized with KOH andprecipitated from acetone to give 8.0 g (100%) of the pure potassiumsalt.

¹H NMR (DMSO-d₆): δ 0.72 (dd, J=5.8, 6.4, 6H), 1.13-1.27 (m, 3H),1.37-1.44 (m 1H), 1.72-1.78 (m, 1H), 1.92-1.98 (m, 1H), 2.67-2.73 (m,2H), 2.80-2.91 (m, 2H), 3.85 (t, J=7.2, 1H), 6.56 (d, J=8.5, 2H), 7.44(d, J=8.5, 2H).

Step C. Preparation of (2S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6-diaminohexanol

A solution of Nα-(4-aminobenzenesulfonyl)-Nα-isobutyl-L-lysine (13.0 g,40.0 mmol, step B) dissolved in MeOH (200 mL) was treated with oftrimethylsilyl chloride (25 mL). The mixture was refluxed 3 h beforestirring at room temperature for 2 h. Afterwards, the solution wasevaporated and placed under high vacuum until a white solid was obtained(14.0 g). This was suspended in dry THF (100 mL) and added dropwise to asolution of LiAlH₄ (5.0 g, 150 mmol) in THF (300 mL). The solution wasstirred for 4 h. After cooling in an ice bath the solution was quenchedby addition of MeOH (50 mL), water (5 mL), then 10% NaOH (5 mL). Thesolvent was evaporated and the product was extracted from theprecipitate with MeOH using a Soxlet apparatus during 18 h. Then, thesolvent was evaporated to form a white solid which was dissolved inEtOH, filtered to eliminate Al₂O₃ and, after cooling, crystallized onstanding (12.0 g, 88%).

¹H NMR (DMSO-d₆): δ 0.82 (m, 6H), 0.97-1.12 (m, 2H), 1.15-1.30 (m, 3H),1.57(m, 1H), 1.84 (m, 1H), 2.40 (t, J=7.0, 2H), 2.75 (m, 1H), 2.85 (m,1H), 3.21 (m, 1H), 3.44 (d, J=6.0, 2H), 5.92(s, 2H), 6.59 (d, J=8.0,2H), 7.39 (d, J=8.0, 2H).

Step D. Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-[N′α-(2-thiophenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6-diaminohexanol (step C)following the indications of general procedure Bd usingNα-(2-thiophenesulfonyl)-L-tryptophan (example 53, step A). Purificationby HPLC gave 200 mg (98%) of the desired material.

¹H NMR (CDCl₃): δ 0.84 (d, J=6.3, 3H), 0.89 (d, J=6.3, 3H), 0.94-1.03(m, 1H), 1.09-1.16 (m, 2H), 1.46-1.49 (m, 1H), 1.93-1.98 (m, 1H),2.82-3.15 (m, 6H), 3.51 (d, J=6.8, 2H), 3.64 (q, J=6.3, 1H), 4.05 (t,J=7.2, 1H), 6.76 (d, J=6.5, 2H), 6.89 (t, J=4.6, 1H), 6.97 (s, 1H), 7.03(t, J=4.5, 1H), 7.19 (t, J=4.5, 1H), 7.23-7.28 (m, 3H), 7.34-7.42 (m,4H), 7.73 (d, J=6.8, 2H).

Example 167 Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N′α-benzenesulfonyl-S-tryptophanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6-diaminohexanol (example166, step C) following the indications of general procedure Bd usingNα-benzenesulfonyl-L-tryptophan (example 4, step A). Purification byHPLC gave 196 mg (96%) of the desired material.

¹H NMR (CDCl₃): δ 0.84 (d J=6.3, 3H), 0.89 (d, J=6.3, 3H), 0.94-1.03 (m,1H), 1.09-1.16 (m, 2H), 1.46-1.49 (m, 1H), 1.93-1.98 (m, 1H), 2.82-3.15(m, 6H), 3.51 (d, J=6.8, 2H), 3.64 (q, J=6.3, 1H), 3.94 (q, J=5.5, 1H),6.76 (d, J=6.5, 2H), 6.95 (t, J=4.5, 1H), 7.19 (t, J=4.5, 1H), 7.23-7.31(m, 6H), 7.42 (t, J=4.5, 1H), 7.60 (d, J=6.8, 2H), 7.73 (d, J=6.8, 2H).

Example 168 Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N′α-acetyl-S-tryptophanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6-diaminohexanol (example166, step C) following the indications of general procedure Bd usingNα-acetyl-L-tryptophan (example 99, step A). Purification by HPLC gave165 mg (96%) of the desired material.

¹H NMR (CDCl₃): δ 0.90-0.98 (m, 6H), 1.09-1.16 (m, 1H), 1.25-1.29 (m,1H), 1.45-1.52 (m, 1H) 1.92-1.97 (m, 1H), 2.0 (s, 3H), 2.38 (s, 3H),2.82-3.10 (m, 4H), 3.11-3.26 (m, 3H), 3.51 (d, J=7.0, 2H), 3.64 (q,J=6.3, 1H), 3.75 (br s, 1H), 6.86 (d, J=5.5, 2H), 6.77 (t, J=5.1, 1H),6.95 (t, J=5.2, 1H), 6.99 (s, 1 H), 7.15 (d, J=6.9, 1H), 7.31 (d, J=7.1,1H), 7.45 (d, J=6.7, 1H).

Example 169 Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N′α-pivaloyl-S-tryptophanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6-diaminohexanol (example166, step C) following the indications of general procedure Bd usingNα-pivaloyl-S-tryptophan (example 100, step A). Purification by HPLCgave 185 mg (97%) of the desired material.

¹H NMR (CDCl₃): δ 0.90-0.98 (m, 6H), 1.09-1.16 (m, 1H), 1.23 (s, 9H),1.25-1.29 (m, 1H), 1.45-1.52 (m, 1H), 1.92-1.97 (m, 1H), 2.82-3.10 (m,4H), 3.11-3.26 (m, 1H), 3.41-3.45 (m, 1H), 4.51 (q, J=5.0, 1H), 4.64 (brs, 1H), 6.86 (d, J=5.5, 2H), 6.77 (t, J=5.1, 1H), 6.85 (t, J=5.2, 1H),6.99 (s, 1H), 7.15 (d, J=6.9, 1H), 7.31 (d, J=7.1, 1H), 7.45 (d, J=6.7,1H).

Example 170 Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-[N′α-(4-morpholinecarbonyl)-S-tryptophanyl]-2,6-diaminohexanol

Step A. Preparation of Nα-(4-morpholinecarbonyl)-L-tryptophan

L-tryptophan was reacted with 4-morpholinecarbonyl chloride under theconditions used in general procedure A giving the title compound as athick oil. This material was used without further purification in thenext step.

Step B. Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-[N′α-(4-morpholinecarbonyl)-S-tryptophanyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6-diaminohexanol (example166, step C) following the indications of general procedure Bd usingNα-(4-morpholinecarbonyl)-L-tryptophan (step A). Purification by HPLCgave 185 mg (97%) of the desired material.

¹H NMR (CDCl₃): δ 0.90-0.98 (m, 6H), 1.09-1.16 (m, 1H), 1.23 (s, 9H),1.25-1.29 (m, 1H), 1.45-1.52 (m, 1H), 1.92-1.97 (m, 1H), 2.82-3.10 (m,4H), 3.11-3.36 (m, 6H), 3.41-3.55 (m, 4H), 4.14 (br s, 1H), 6.86 (d,J=5.5, 2H), 6.77 (t, J=5.1, 1H), 6.85 (t, J=5.2, 1H), 6.99 (s, 1H), 7.15(d, J=6.9, 1H), 7.31 (d, J=7.1, 1H), 7.45 (d, J=6.7, 1H).

Example 171 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-(4-morpholinecarbonyl)-S-tryptophanyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-(4-morpholinecarbonyl)-L-tryptophan (example 170, step A).Purification by HPLC gave 110 mg (57%) of the desired material.

¹H NMR (CDCl₃): δ 0.90-0.98 (m, 6H), 1.09-1.16 (m, 1H), 1.23 (s, 9H),1.25-1.29 (m, 1H), 1.45-1.52 (m, 1H), 1.92-1.97 (m, 1H), 2.33 (s, 3H),2.82-3.10 (m, 4H), 3.11-3.36 (m, 6H), 3.41-3.55 (m, 4H), 4.14 (br s,1H), 6.86 (d, J=5.5, 2H), 6.77 (t, J=5.1, 1H), 6.85 (t, J=5.2, 1H), 6.99(s, 1H), 7.15 (d, J=6.9, 1H), 7.31 (d, J=7.1, 1H), 7.45 (d, J=6.7, 1H).

Example 172 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methoxybenzenesulfonyl)-6-N-(N′α-acetyl-S-tryptophanyl)-2,6-diaminohexanol

Treatment of (2S,2′S)2-N-isobutyl-2-N-(4-methoxybenzenesulfonyl)-6-N-(S-tryptophanyl)-2,6-diaminohexanoltrifluoroacetic acid salt (example 146) with acetyl chloride, usingsimilar reaction conditions as for example 65 (step B) which uses4-methylbenzenesulfonyl chloride, afforded the desired product in 86%yield.

¹H NMR (DMSO-d₆): δ 0.82 (t, J=7.0, 6H), 1.02 (s, 9H), 1.00-1.30 (m,5H), 1.77 (s, 3H), 2.80-2.95 (m, 5H), 3.03 (dd, J=13.5, 7.0, 1H), 3.77(m, 1H), 3.81 (s, 3H), 3.90 (m, 1H), 3.98 (m, 1H), 4.45 (m, 1H), 6.96(t, J=7.5, 1H), 7.04 (t, J=7.5, 1H), 7.10 (d, J=8.0, 2H), 7.30 (d,J=7.5, 1H), 7.58 (d, J=7.5, 1H), 7.70 (d, J=8.6, 2H), 7.85 (t, J=5.5,1H), 7.99 (d, J=8.5, 1H), 10.75 (s, 1H).

Example 173 Preparation of (2S,2′S)2-N-cyclopentylmethyl-2-N-(4-methoxybenzenesulfonyl)-6-N-(N′α-acetyl-S-tryptophanyl)-2,6-diaminohexanol

Treatment of (2S,2′S)2-N-cyclopentylmethyl-2-N-(4-methoxybenzenesulfonyl)-6-N-(S-tryptophanyl)-2,6-diaminohexanoltrifluoroacetic acid salt (example 152) with acetyl chloride, usingsimilar reaction conditions as for example 65 (step B) which uses4-methylbenzenesulfonyl chloride, afforded the desired product in 76%yield.

¹H NMR (DMSO-d₆): δ 0.93-1.30 (m, 7H), 1.40-1.68 (m, 7H), 1.77 (s, 3H),2.18 (m, 1H), 2.84-3.10 (m, 5H), 3.30 (m, 2H), 3.50 (m, 1H), 3.81 (s,3H), 4.45 (m, 1H), 4.65 (m, 1H), 6.95-7.13 (m, 5H), 7.30 (d, J=8.3, 1H),7.56 (d, J=8.5, 1H), 7.71 (d, J=8.2, 2H), 7.85 (t, J=5.5, 1H), 7.98 (d,J=8.0, 1H), 10.75 (s, 1H).

Example 174 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methoxybenzyl)-N′α-phenylglycyl]-L-lysine

Step A. Preparation of Nα-(4-methoxybenzyl)-Nα-phenylglycine

The title compound was prepared from N-phenylglycine (1.51 g, 10.0 mmol)as described for the preparation of N-benzyl-N-phenylglycine (example114, step A) using 4-methoxybenzylchloride (1.72 g, 11.0 mmol) insteadof benzylbromide. The crude material was purified by flashchromatography using 9:1 CH₂Cl₂/hexane. The product was isolated as ayellow solid (2.33 g, 86% yield).

¹H NMR (DMSO-d₆): δ 3.72 (s, 3H), 4.11 (s, 2H), 4.51 (s, 2H), 6.59 (m,3H), 6.87 (d, J=7.5, 2H), 7.11 (t, J=7.9, 2H), 7.21 (d, J=8.2, 2H), 12.6(s, 1H).

Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methoxybenzyl)-N′α-phenylglycyl]-L-lysine

The title product was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine hydrochloride (360 mg,1.0 mmol, example 1, step E) following the indications of generalprocedure Ba using Nα-(4-methoxybenzyl)-Nα-phenylglycine (271 mg, 1.0mmol, step A) and N,N-carbonyldiimidazole (195 mg, 1.2 mmol). The crudematerial was purified by preparative HPLC. The product was isolated as asolid (39 mg, 6% yield).

¹H NMR (DMSO-d₆): δ 0.81 (t, J=6.9, 6H), 1.17 (m, 2H), 1.33 (m, 2H),1.48 (m, 1H), 1.80 (m, 1H), 1.90 (m, 1H), 2.37 (s, 3H), 2.85-3.05 (m,4H), 3.71 (s, 3H), 3.90 (s, 2H), 4.26 (t, J=7.2, 1H), 4.54 (s, 2H), 6.61(t, J=8.7, 3H), 6.87 (d, J=8.4, 2H), 7.11 (t, J=7.4, 2H), 7.17 (d,J=8.6, 2H), 7.36 (d, J=7.6, 2H), 7.66 (d, J=8.3, 2H), 7.83 (t, J=5.1,1H), 12.75 (s, 1H). LC-MS: 610 (M+H)⁺, 99% pure.

Example 175 Preparation of (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N′α-(4-methoxybenzyl)-N′α-phenylglycyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (345 mg,1.0 mmol, example 116, step A) following the indications of generalprocedure Ba using Nα-(4-methoxybenzyl)-Nα-phenylglycine (271 mg, 1.0mmol, example 174, step A) and N,N-carbonyldiimidazole (195 mg, 1.2mmol). The crude material was purified by preparative HPLC. The productwas isolated as a yellow solid (105 mg, 18% yield).

¹H NMR (DMSO-d₆): 67 0.82 (m, 6H), 1.01 (m, 2H), 1.23 (m, 3H), 1.48 (m,1H), 1.86 (m, 1H), 2.36 (s, 3H), 2.85 (m, 1H), 2.95 (m, 3H), 3.26 (dd,J=11.2, 6.3, 1Ha), 3.32 (dd, J=11.2, 6.3, 1Hb), 3.52 (m, 1H), 3.71 (s,3H), 3.89 (s, 2H), 4.35 (m, 1H), 4.54 (s, 2H), 6.61 (m, 3H), 6.86 (d,J=8.6, 2H), 7.11 (t, J=8.0, 2H), 7.17 (d, J=7.9, 2H), 7.36 (t, J=7.0,2H), 7.66 (d, J=8.5, 2H), 7.78 (t, J=4.9, 1H). LC-MS: 596 (M+H)⁺, 95%pure.

Example 76 Preparation of (2S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-[N′α-(4-methoxybenzyl)-N′α-phenylglycyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6-diaminohexanol (350 mg,1.0 mmol example 166, step C) following the indications of generalprocedure Ba using Nα-(4-methoxybenzyl)-Nα-phenylglycine (271 mg, 1.0mmol, example 174, step A) and N,N-carbonyldiimidazole (195 mg, 1.2mmol). The crude material was purified by preparative HPLC. The productwas isolated as a yellow solid (87 mg, 14% yield).

¹H NMR (DMSO-d₆): δ 0.81 (t, J=7.0, 6H), 1.10 (m, 2H), 1.27 (m, 3H),1.51 (m, 1H), 1.85 (m, 1H), 2.75 (dd, J=14.1, 7.3, 1Ha), 2.83 (dd,J=14.1, 7.3, 1Hb), 2.98 (m, 2H), 3.22 (m, 1H), 3.30 (m, 1H), 3.45 (m,1H), 3.71 (s, 3H), 3.90 (s, 2H), 4.54 (s, 2H), 6.60 (m, 5H), 6.86 (d,J=8.8, 2H), 7.11 (t, J=7.6, 2H), 7.17 (d, J=7.7, 2H), 7.39 (d, J=8.7,2H), 7.80 (t, J≅5.0, 1H). LC-MS: 597 (M+H)⁺, 98% pure.

Example 177 Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N′α-methoxycarbonyl-S-phenylalanyl)-2,6-diamninohexanol

Step A. Preparation of Nα-methoxycarbonyl-L-phenylalanine

L-phenylalanine was reacted with methyl chloroformate under theconditions used in general procedure A giving the title compound as anoil which was used without further purification in the next step.

Step B. Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N′α-methoxycarbonyl-S-phenylalanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6-diaminohexanol (example166, step C) following the indications of general procedure Bd usingNα-methoxycarbonyl-L-phenylalanine (step A). Purification by HPLC gave159 mg (98%) of the desired material.

¹H NMR (CDCl₃): δ 0.79 (d, J=6.3, 3H), 0.82 (d, J=6.3, 3H), 1.08-1.11(m, 2H), 1.23-1.31 (m, 2H), 1.45-1.52 (m, 1H), 1.92-1.97 (m, 1H), 1.98(d, J=9.0, 3H), 2.84-3.09 (m, 6H), 3.51 (d, J=7.1, 2H), 3.59 (s, 3H),3.61-3.64 (m, 1H), 4.51 (q, J=6.9, 1H), 6.76 (d, J=8.1, 2H), 7.09-7.26(m, 5H), 7.73 (d, J=8.1, 2H). LC-MS: 549.7 (M+H)⁺, 98% pure.

Example 178 Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N′α-methoxycarbonyl-S-tryptophanyl)-2,6-diaminohexanol

Step A. Preparation of Nα-methoxycarbonyl-L-tryptophan

L-tryptophan was reacted with methyl chloroformate under the conditionsused in general procedure A giving the title compound as an oil whichwas used without further purification in the next step.

Step B. Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N′α-methoxycarbonyl-S-tryptophanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6-diaminohexanol (example166, step C) following the indications of general procedure Bd usingNα-methoxycarbonyl-L-tryptophan (step A). Purification by HPLC gave 155mg (87%) of the desired material.

¹H NMR (CDCl₃): δ 0.90-0.98 (m, 6H), 1.09-1.16 (m, 1H), 1.25-1.29 (m,1H), 1.45-1.52 (m, 1H), 1.92-1.97 (m, 1H), 2.82-3.10 (m, 4H), 3.11-3.26(m, 3H), 3.51 (d, J=7.0, 2H), 3.59 (s, 3H), 3.64 (q, J=6.3, 1H), 3.75(br s, 1H), 6.86 (d, J=5.5, 2H), 6.77 (t, J=5.1, 1H), 6.95 (t, J=5.2,1H), 7.15 (d, J=6.9, 1H), 7.31 (d, J=7.1, 1H), 7.45(d, J=6.7, 1H).LC-MS: 588.7 (M+H)⁺, 98% pure.

Example 179 Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-[N′α-(4-nitrobenzenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6-diaminohexanol (example166, step C) following the indications of general procedure Bd usingNα-(4-nitrobenzenesulfonyl)-L-tryptophan (example 5, step A).Purification by HPLC gave 255 mg (60%) of the desired material.

LC-MS: 715.8 (M+H)⁺, 98% pure.

Example 180 Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-[N′α-(4-morpholinecarbonyl)-S-phenylalanyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6-diaminohexanol (example166, step C) following the indications of general procedure Bd usingNα-(4-morpholinecarbonyl)-L-phenylalanine (example 51, step A).Purification by HPLC gave 165 mg (91%) of the final product.

¹H NMR (CDCl₃): δ 0.79 (d, J=6.3, 3H), 0.82 (d, J=6.3, 3H), 1.08-1.11(m, 2H), 1.23-1.31 (m, 2H), 1.45-1.52 (m, 1H), 1.92-1.97 (m, 1H),2.84-3.09 (m, 6H), 3.21-3.31 (m, 4H), 3.45-3.64 (m, 7H), 4.48 (br s,1H), 6.88 (d, J=8.1, 2H), 7.09-7.26 (m, 5H), 7.43 (d, J=8.1, 2H). LC-MS:604.8 (M+H)⁺, 98% pure.

Example 181 Preparation of (2S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N′α-isobutyl-N′α-phenylglycyl)-2,6-diaminohexanol

Step A. Preparation of N-isobutyl-N-phenylglycine

The title compound was prepared from N-phenylglycine (1.0 g, 5.5 mmol)as described for the preparation of N-benzyl-N-phenylglycine (example114, step A) using isobutyl iodide (2.0 mL, 17.3 mmol) instead ofbenzylbromide. The crude material was purified by flash chromatographyusing hexane/EtOAc/CHCl₃ (7:3:2) as the eluent. The product was isolatedas a brown oil (560 mg, 41% yield).

¹H NMR (CDCl₃): δ 0.94 (m, 6H), 2.03 (m, 1H), 3.16 (d, J=7.2, 2H), 4.06(s, 2H), 6.66 (d, J=8.2, 2H), 6.75 (t, J=7.2, 1H), 7.21 (t, J=8.0, 2H).

Step B. Preparation of (2S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N′α-isobutyl-N′α-phenylglycyl)-2,6-diaminohexanol

A solution of N-isobutyl-N-phenylglycine (200 mg, 1.0 mmol, step A) inDMF (5.0 mL) was treated with EDC (290 mg, 1.5 mmol) and HOBt (100 mg,0.75 mmol) for a period of 10 min under an inert atmosphere or argon.Then, (2S) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6-diaminohexanol(345 mg, 1.0 mmol example 166, step C) was added and the reactionmixture was stirred for a period of 6 h. Afterwards, a 10% aqueouscitric acid solution was added and the product extracted with EtOAc (10mL, 3×). The organic phase was washed with water 10 mL (3×) and withbrine (5 mL). The organic phase was dried with MgSO₄, filtered andevaporated to a solid. The crude material was purified by preparativeHPLC. The product was isolated as a yellow solid (125 mg, 24% yield).

¹H NMR (DMSO-d₆): δ 0.81 (t, J=6.5, 6H), 0.88 (d, J=8.1, 6H), 1.05 (m,1H), 1.13 (m, 1H), 1.26 (m, 3H), 1.55 (m, 1H), 1.84 (m, 1H), 1.98 (m,1H), 2.74 (dd, J=13.6, 7.1, 1H), 2.83 (dd, J=14.2, 7.5, 1H), 2.95 (d,J=6.4, 2H), 3.18 (d, J=7.3, 2H), 3.23 (m, 1H), 3,44 (m, 1H), 3.85 (s,2H), 4.59 (t, J=5.0, 1H), 5.90 (s, 2H), 6.58 (m, 5H), 7.11 (t, J=7.4,2H), 7.38 (d, J=8.6, 2H), 7.67 (t, J=5.0, 1H). LC-MS: 553 (M+H)⁺, 98%pure.

Example 182 Preparation of (2S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N′α-benzyl-N′α-phenylglycyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6-diaminohexanol (345 mg,1.0 mmol, example 166, step C) as described above for the preparation of2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N′α-isobutyl-N′α-phenylglycyl)-2,6-diaminohexanol(example 183) using N-benzyl-N-phenylglycine (241 mg, 1.0 mmol, example114, step A) instead of N-isobutyl-N-phenylglycine. The crude materialwas purified by preparative HPLC. The product was isolated as a yellowsolid (370 mg, 65% yield).

¹H NMR (DMSO-d₆): δ 0.81 (t, J=6.9, 6H), 1.05 (m, 1H), 1.13 (m, 1H),1.28 (m, 3H), 1.55 (m, 1H), 1.84 (m, 1H), 2.74 (dd, J=12.6, 7.0, 1H),2.85 (dd, J=11.3, 5.9, 1H), 2.98 (d, J=6.0, 2H), 3.24 (dd, J=11.7, 6.3,1H), 3.30 (dd, J=8.9, 5.0, 1H), 3.45 (m, 1H), 3.94 (s, 2H), 4.62 (s,2H), 5.35 (br s, 2H), 6.60 (m, 5H), 7.11 (t, J=7.7, 2H), 7.24 (m, 3H),7.31 (t, J=7.5, 2H), 7.39 (d, J=7.6, 2H), 7.83 (t, J=5.1, 1H). LC-MS:567 (M+H)⁺, 99% pure.

Example 183 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(S-tryptophanyl)-2,6-diaminohexanoltrifluoroacetic acid salt

Step A. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-L-tryptophanyl)-L-lysinemethyl ester

To a stirred solution ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine methyl ester (370 mg,1 mmol, example 65, step C) in THF/K₂CO₃ (1M) (3 mL/3 mL) was addedNα-tert-butoxycarbonyl-L-tryptophan N-hydroxysuccinimide ester (550 mg,1.2 mmol). The reaction mixture was stirred overnight then diluted with1N HCl and extracted with EtOAc. The organic layer was dried (MgSO₄) andconcentrated. The crude was purified by flash chromatography usinghexane/EtOAc as eluent to afford the desired product (85% yield).

¹H NMR (DMSO-d₆): δ 0.81 (t, J=7.0, 6H), 1.10-1.50 (m, 5H), 1.31 (s,9H), 1.75-1.90 (m, 2H), 2.38 (s, 3H), 2.82-3.10 (m, 5H), 3.44 (s, 3H),4.12 (m, 1H), 4.30 (t, J=5.0, 1H), 6.67 (d, J=8.2, 1H), 6.90-7.12 (m,5H), 7.30 (d, J=8.0, 1H), 7.40 (d, J=7.5, 2H), 7.55 (m, 1H), 7.65 (d,J=7.5, 2H), 7.78 (m, 1H), 10.77 (s, 1H).

Step B. Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-tert-butoxycarbonyl-S-tryptophanyl)-2,6-diaminohexanol

Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-tert-butoxycarbonyl-L-tryptophanyl)-L-lysinemethyl ester (step A) was reduced with LiAlH₄ following the indicationsof general procedure G. The final product was obtained with 78% yield.

¹H NMR (DMSO-d₆): δ 0.84 (t, J=7.0, 6H), 0.90-1.27 (m, 5H), 1.31 (s,9H), 1.52 (m, 1H), 1.85 (m, 1H), 2.37 (s, 3H), 2.80 and 3.02 (ABX,J=14.0, 7.2, 2H), 2.90 (m, 2H), 3.33 (m, 1H), 3.50 (m, 1H), 4.15 (m,1H), 4.65 (t, J=5.0, 1H), 6.65 (d, J=7.8, 1H), 6.92-7.12 (m, 5H), 7.30(d, J=7.8, 1H), 7.35 (d, J=7.5, 2H), 7.57 (d, J=8.0, 1H), 7.67 (d,J=7.7, 2H), 7.73 (t, J=5.0, 1H), 10.77 (s, 1H).

Step C. Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(S-tryptophanyl)-2,6-diaminohexanoltrifluoroacetic acid salt

This product was obtained quantitatively by treating (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-tert-butoxycarbonyl-S-tryptophanyl)-2,6-diaminohexanol(step B) with TFA in CH₂Cl₂. 1H NMR (DMSO-d₆): δ 0.80 (d, J=7.0, 3H),0.83 (d, J=7.0, 3H), 0.92-1.60 (m, 6H), 1.88 (m, 1H), 2.37 (s, 3H),2.78-3.00 (m, 4H), 3.05 and 3.20 (ABX, J=14.0, 7.0, 2H), 3.22 and 3.30(ABX, J=14.2, 7.0, 2H), 3.50 (m, 1H), 3.86-3.97 (m, 2H), 4.35 (m, 1H),6.95-7.12 (m, 1H), 7.18 (s, 1H), 7.38 (d, J=8.2, 1H), 7.60 (m, 1H), 7.70(d, J=8.2, 2H), 8.05 (br s, 3H), 8.34 (m, 1H), 10.99 (s, 1H).

Example 184 Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N′α-isobutyryl-S-tryptophanyl)-2,6-diaminohexanol

Step A. Preparation of Nα-isobutyryl-L-tryptophan

L-tryptophan was reacted with isobutyryl chloride under the conditionsused in general procedure A giving the title compound which wasrecrystallised from DCM (55%). This material was used without furtherpurification in the next step.

Step B. Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N′α-isobutyryl-S-tryptophanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6-diaminohexanol (example166, step C) following the indications of general procedure Bd usingNα-isobutyryl-L-tryptophan (step A). Purification by HPLC gave 121 mg(67%) of the desired material.

¹H NMR (DMSO-d₆): δ 0.80-0.90 (m, 9H), 0.95-0.97 (d, J=6.6, 3H),1.09-1.16 (m, 1H), 1.25-1.29 (m, 1H), 1.45-1.52 (m, 1H), 1.92-1.97 (m,1H), 2.33-2.45 (m, 1H), 2.82-2.99 (m, 4H), 3.26-3.33 (m, 1H), 3.51-3.55(m, 1H), 4.64 (br s, 1H), 6.66 (d, J=5.5, 2H), 6.87 (t, J=5.1, 1H), 6.95(t, J=5.2, 1H), 6.99 (s, 1 H), 7.15 (d, J=6.9, 1H), 7.31 (d, J=7.1, 1H),7.45 (d, J=6.7, 1H). LC-MS: 600.8 (M+H)⁺, 99% pure.

Example 185 Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N′α-tert-butylacetyl-S-tryptophanyl)-2,6-diaminohexanol

Step A. Preparation of Nα-tert-butylacetyl-L-tryptophan

L-tryptophan was reacted with tert-butylacetyl chloride under theconditions used in general procedure A giving the title compound whichwas recrystallised from DCM (81%). This material was used as such in thenext step.

Step B. Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N′α-tert-butylacetyl-S-tryptophanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6-diaminohexanol (example166, step C) following the indications of general procedure Bd usingNα-tert-butylacetyl-L-tryptophan (step A). Purification by HPLC gave 133mg (70%) of the desired material.

¹H NMR (DMSO-d₆): δ 0.80-0.90 (m, 15H), 1.09-1.16 (m, 1H), 1.25-1.29 (m,1H), 1.45-1.52 (m, 1H), 1.82-1.87 (m, 1H), 1.97 (s, 2H), 2.33-2.45 (m,1H), 2.82-2.99 (m, 4H), 3.26-3.33 (m, 1H), 3.51-3.55 (m, 1H), 4.64 (brs, 1H), 6.66 (d, J=5.5, 2H), 6.87 (t, J=5.1, 1H), 6.95 (t, J=5.2, 1H),6.99 (s, 1H), 7.15 (d, J=6.9, 1H), 7.31 (d, J=7.1, 1H), 7.45 (d, J=6.7,1H). LC-MS: 628.8 (M+H)⁺, 98% pure.

Example 186 Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N′α-benzoyl-S-tryptophanyl)-2,6-diaminohexanol

Step A. Preparation of Nα-benzoyl-L-tryptophan

L-tryptophan was reacted with benzoyl chloride under the conditions usedin general procedure A giving the title compound which wasrecrystallised from DCM (51%). This compound was used without furtherpurification in the next step.

Step B. Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N′α-benzoyl-S-tryptophanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6-diaminohexanol (example166, step C) following the indications of general procedure Bd usingNα-benzoyl-L-tryptophan (step A). Purification by HPLC gave 122 mg (64%)of the desired material.

¹H NMR (CDCl₃): δ 0.84(d, J=6.3, 3H), 0.89 (d, J=6.3, 3H), 0.94-1.03 (m,1H), 1.09-1.16 (m, 2H), 1.46-1.49 (m, 1H), 1.93-1.98 (m, 1H), 2.39 (s,3H), 2.82-3.15 (m, 6H), 3.51 (d, J=6.8, 2H), 3.64 (q, J=6.3, 1H), 3.94(q, J=5.5, 1H), 6.95 (t, J=4.5, 1H), 7.19 (t, J=4.5, 1H), 7.23-7.31 (m,6H), 7.42 (t, J=4.5, 1H), 7.60 (d, J=6.8, 2H), 7.73 (d, J=6.8, 2H).LC-MS: 634.8 (M+H)⁺, 98% pure.

Example 187 Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-[′α-(4-aminobenzenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanol

The title compound was obtained from (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-[N′α-(4-nitrobenzenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanol(200 mg, 0.28 mmol, example 179) by catalytic hydrogenation followingthe conditions of general procedure E. Purification by HPLC gave 191 mg(99%) of the desired material.

¹H NMR (DMSO-d₆): δ 0.73 (d, J=6.3, 3H), 0.75 (d, J=6.3, 3H), 1.00-1.11(m, 4H), 1.32-1.35 (m, 1H), 1.66-1.69 (m, 1H), 1.83-1.88 (m, 1H), 2.48(br s, 6H), 2.59-2.67 (m, 2H), 2.84-2.96 (m, 2H), 3.20 (d, J=6.5, 2H),4.21 (t, J=7.2, 1H), 6.66 (d, J=7.1, 2H), 6.72 (d, J=7.1, 2H), 6.85 (t,J=4.0, 1H), 7.09 (t, J=4.0, 2H), 7.28 (d, J=7.1, 1H), 7.33 (d, J=7.1,2H), 7.60 (t, J=4.0, 1H). LC-MS: 685.8 (M+H)⁺, 98% pure.

Example 188 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-benzenesulfonyl-S-cyanoalanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-benzenesulfonyl-L-asparagine (example 25, step A). In this particularreaction an excess EDC (2.5 eq.) was used which dehydrated the amidefunction on the asparagine moiety. Purification by HPLC gave 6 mg (9%)of the desired material.

LC-MS: 579.2 (M+H)⁺, 92% pure.

Example 189 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzenesulfonyl-L-tryptophanyl)-L-lysineamide

A solution ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-benzenesulfonyl-L-tryptophanyl)-L-lysine(100 mg, 0.15 mmol, example 4) dissolved in EtOAc (3 mL) was treatedwith DCC (30 mg, 0.15 mmol) and N-hydroxysuccinimide (17 mg, 0.15 mmol).The resulting solution was stirred overnight at room temperature. Then,the reaction mixture was filtered through celite. The organic solventwas evaporated and the crude residue dissolved in THF (5 mL) was treatedwith concentrated NH₄OH (1 mL). The reaction mixture was left in therefrigerator overnight. Afterwards, it was filtered through celite andthe solvent was evaporated to give 98 mg (97%) of the desired titlecompound after purification by HPLC.

LC-MS: 682.8 (M+H)⁺, 99% pure.

Example 190 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-methoxycarbonyl-S-tryptophanyl)-2,6-diaminohexanol

The title product was prepared from (28)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-methoxycarbonyl-L-tryptophan (example 178, step A). Purification byHPLC gave 21 mg (15%) of the desired material.

¹H NMR (CDCl₃): δ 0.90-0.98 (m, 6H), 1.09-1.16 (m, 1H), 1.25-1.29 (m,1H), 1.45-1.52 (m, 1H), 1.92-1.97 (m, 1H), 2.38 (s, 3H), 2.82-3.10 (m,4H), 3.11-3.26 (m, 3H), 3.51 (d, J=7.0, 2H), 3.59 (s, 3H), 3.64 (q,J=6.3, 1H), 3.75 (br s, 1H), 6.77 (t, J=5.1, 1H), 6.95 (t, J=5.2, 1H),6.99 (s, 1H), 7.15-7.25 (m, 3H), 7.31 (d, J=7.1, 1H), 7.75 (d, J=6.7,1H). LC-MS: 587 (M+H)⁺, 99% pure.

Example 191 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-benzoyl-S-tryptophanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-benzoyl-L-tryptophan (example 186, step A). Purification by HPLC gave41 mg (28%) of the desired material.

¹H NMR (CDCl₃): δ 0.90-0.98 (m, 6H), 1.09-1.16 (m, 1H), 1.25-1.29 (m,1H), 1.45-1.52 (m, 1H), 1.92-1.97 (m, 1H), 2.82-3.10 (m, 4H), 3.11-3.26(m, 3H), 3.51 (d, J=7.0, 2H), 3.59 (s, 3H), 3.64 (q, J=6.3, 1H), 4.08(br s, 1H), 6.86 (d, J=5.5, 2H), 6.77 (t, J=5.1, 1H), 6.95 (t, J=5.2,1H), 6.99 (s, 1H), 7.15 (d, J=6.9, 1H), 7.31 (d, J=7.1, 1H), 7.45 (d,J=6.7, 1H). LC-MS: 633.8 (M+H)⁺, 99% pure.

Example 192 Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-isobutoxycarbonyl-S-tryptophanyl)-2,6-diaminohexanol

Step A. Preparation of Nα-isobutoxycarbonyl-L-tryptophan

L-tryptophan was reacted with isobutyl chloroformate under theconditions used in general procedure A giving the title compound whichwas recrystallised neat. This product was used without furtherpurification in the next step.

Step B. Preparation of (2S,2′S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N′α-isobutoxycarbonyl-S-tryptophanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6-diaminohexanol (example116, step A) following the indications of general procedure Bd usingNα-isobutoxycarbonyl-L-tryptophan (step A). Purification by HPLC gave 64mg (40%) of the desired material.

¹H NMR (DMSO-d₆): δ 0.80-0.91 (m, 9H), 0.94-0.97 (d, J=6.6, 3H),1.09-1.16(m, 1H), 1.25-1.29 (m, 1H), 1.45-1.52 (m, 1H), 1.92-1.97 (m,1H), 2.33-2.45 (m and s, 4H), 2.82-2.99 (m, 6H), 3.26-3.33 (m, 1H),3.51-3.55 (m, 1H), 4.44 (br s, 1H), 6.87 (t, J=5.1, 1H), 6.95 (t, J=5.2,1H), 6.99 (s, 1H), 7.15-7.22 (m, 3H), 7.31 (d, J=7.1, 1H), 7.65 (d,J=6.7, 1H). LC-MS: +H)⁺, 99% pure.

Example 193 Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N′α-isobutoxycarbonyl-S-tryptophanyl)-2,6-diaminohexanol

The title product was prepared from (2S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6-diaminohexanol (example166, step C) following the indications of general procedure Bd usingNα-isobutoxycarbonyl-L-tryptophan (example 192, step A). Purification byHPLC gave 151 mg (95%) of the final product.

¹H NMR (DMSO-d₆): δ 0.80-0.90 (m, 9H), 0.95-0.97 (d, J=6.6, 3H),1.09-1.16 (m, 1H), 1.25-1.29 (m, 1H), 1.45-1.52 (m, 1H), 1.92-1.97 (m,1H), 2.33-2.45 (m, 1H), 2.82-2.99 (m, 6H), 3.26-3.33 (m, 1H), 3.51-3.55(m, 1H), 4.44 (br s, 1H), 6.66 (d, J=5.5, 2H), 6.87 (t, J=5.1, 1H), 6.95(t, J=5.2, 1H), 6.99 (s, 1 H), 7.15 (d, J=6.9, 1H), 7.31 (d, J=7.1, 1H),7.45 (d, J=6.7, 1H). LC-MS: 630.2 (M+H)⁺, 99% pure.

Example 194 Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-[N′α-(4-nitrobenzenesulfonyl)-S-phenylalanyl]-2,6-diaminohexanol

The title product was prepared from (2S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6-diaminohexanol (example166, step C) following the indications of general procedure Bd usingNα-(4-nitrobenzenesulfonyl)-L-phenylalanine (example 10, step A).Purification by HPLC of about half (50 mg) of the crude material gave 20mg (40%) of the final product.

LC-MS: 676.8 (M+H)⁺, 95% pure.

Example 195 Preparation of (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-[N′α-(4-aminobenzenesulfonyl)-S-phenylalanyl]-2,6-diaminohexanol

The title compound was obtained from (2S,2′S)2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-[N′α-(4-nitrobenzenesulfonyl)-S-phenylalanyl]-2,6-diaminohexanol(50 mg, 0.07 mmol, example 194) by catalytic hydrogenation following theconditions of general procedure E. Purification by HPLC gave 31 mg (77%)of the desired material.

LC-MS: 646.8 (M+H)⁺, 99% pure.

Example 196 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methoxyanilinecarbonyl)-L-phenylalanyl]-L-lysine

The title compound was prepared from solid phase boundNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(9-fluorenylmethoxycarbonyl)-L-lysine(450 mg, 0.125 mmol) as described in general procedure Bb usingcommercially available Nα-(9-fluorenylmethoxycarbonyl)-L-phenylalanine(400 mg, 0.9 mmol). After the coupling reaction, the resin was once moredeprotected and activated with N,N-carbonyldiimidazole (5-10 foldexcess) for 30 min. after which time the resin was washed with DCM (4×)before 4-methoxyaniline was added. The tube was sealed and left for aperiod of 12 h. Afterwards, the product was cleaved from the resin usingTFA as indicated in general procedure Bb. The final product was purifiedby preparative HPLC to yield 11 mg (13%) of the desired material.

¹H NMR (CDCl₃): δ 0.83 (d, J=6.9, 6H), 1.08-1.11 (m, 2H), 1.33-1.55 (m,2H), 1.45-1.52 (m, 1H), 1.79-1.89 (m, 2H), 2.36 (s, 3H), 2.85-3.27 (m,6H), 3.55 (s, 2H), 3.79 (s, 3H), 4.21 (s, 2H), 4.33 (t, J=4.5, 1H), 6.69(d, J=8.2, 2H), 6.99-7.19 (m, 3H), 7.15-7.26 (m, 7H), 7.73 (d, J=8.1,2H). LC-MS: 651.8 (M−H)⁻, 99% pure.

Example 197 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-pyrrolidinecarbonyl-L-phenylalanyl)-L-lysine

This compound was prepared as described for the preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methoxyanilinecarbonyl)-L-phenylalanyl]-L-lysine(example 196) using pyrrolidine instead of 4-methoxyaniline. The crudematerial was purified by preparative HPLC to give 15 mg, 20% of thedesired material.

LC-MS: 599.7 (M−H)⁻, 99% pure.

Example 198 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-methylaminocarbonyl-L-phenylalanyl)-L-lysine

This compound was prepared as described for the preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methoxyanilinecarbonyl)-L-phenylalanyl]-L-lysine(example 196) using methylamine instead of 4-methoxyaniline. The crudematerial was purified by preparative HPLC to give 4 mg, 6% of thedesired material.

¹H NMR (CDCl₃): δ 0.83 (d, J=6.9, 6H), 1.08-1.11 (m, 2H), 1.23-1.25 (m,2H), 1.45-1.52 (m, 1H), 1.89-1.99 (m, 2H), 2.32 (s, 3H), 2.70 (s, 3H),2.94-3.09 (m, 6H), 4.23 (t, J=5.9, 1H), 4.61 (m, 1H), 7.09-7.26 (m, 7H),7.73 (d, J=8.1, 2H). LC-MS: 559.2 (M−H)⁻, 99% pure.

Example 199 Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-(N′α-ethanolaminocarbonyl-L-phenylalanyl)-L-lysine

This compound was prepared as described for the preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Nε-[N′α-(4-methoxyanilinecarbonyl)-L-phenylalanyl]-L-lysine(example 196) using ethanolamine instead of 4-methoxyaniline. The crudematerial was purified by preparative HPLC to give 4.1 mg, 5% of thedesired material.

¹H NMR (CDCl₃): δ 0.79 (d, J=6.3, 3H), 0.82 (d, J=6.3, 3H), 1.08-1.11(m, 2H), 1.23-1.31 (m, 2H), 1.45-1.52 (m, 1H), 1.92-1.97 (m, 1H), 2.39(s, 3H), 2.84-3.29 (m, 8H), 3.45-3.64 (m, 2H), 4.11 (br s, 1H), 4.48 (brs, 1H), 7.09-7.26 (m, 7H), 7.73 (d, J=8.1, 2H). LC-MS: 589.7 (M−H)⁻, 99%pure.

Example 200 Preparation ofNα-benzenesulfonyl-Nα-isobutyl-Nε-[N′α-(4-methylbenzenesulfonyl)-L-phenylalanyl]-L-lysine

Step A. Preparation ofNα-benzenesulfonyl-Nα-isobutyl-L-α-amino-ε-caprolactam

Nα-isobutyl-L-α-amino-ε-caprolactam (example 1, step C) (4.0 g, 21.3mmol, free base) was dissolved in DCM (100 mL) and treated withdiisopropylethylamine (4.0 mL) followed by freshly recrystallizedbenzenesulfonyl chloride (3.5 g, 21 mmol). The mixture was stirredovernight (TLC shows the reaction to be complete after 2 h). Thesolution was extracted with 1N HCl and the organic layer was dried andevaporated to give 5.5 g (80%) of pure product. This compound was usedwithout further purification in the next step.

Step B. Preparation of Nα-benzenesulfonyl-Nα-isobutyl-L-lysine

A mixture of Nα-benzenesulfonyl-Nα-isobutyl-L-α-amino-ε-caprolactam (5.0g, 15 mmol) and 6N HCl (50 mL) was refluxed for 6 h until all solids haddisappeared. Afterwards, the solution was evaporated and the resultingsolid was triturated with THF to give 5.2 g, 96% of the desiredmaterial. LC-MS: 346 (M+H)⁺, 99% pure.

Step C. Preparation ofNα-benzenesulfonyl-Nα-isobutyl-Nε-[Nα′-(4-methylbenzenesulfonyl)-L-phenylalanyl]-L-lysine

A suspension of Nα-benzenesulfonyl-Nα-isobutyl-L-lysine (150 mg, 0.5mmol), in THF (10 mL) was treated with a 1N NaOH (3.0 mL) to pH 10. Asolution of commercially availableNα-(4-methylbenzenesulfonyl)-L-phenylalanine acid chloride (187 mg, 0.5mmol), in dry THF (10 mL) was added to the suspension and stirred for 4h. Afterwards, water (2 mL) was added resulting in a clear solution.Then, EtOAc (30 mL) was added and the organic phase was washed with 1NHCl. The organic phase was removed. Evaporation of the solvent gave acrude product which was purified by preparative HPLC to yield 19 mg (6%)of the title compound.

¹H NMR (CDCl₃): δ 0.76 (d, J=6.3, 3H), 0.80 (d, J=6.3, 3H), 1.10-1.20(m, 2H), 1.26-1.33 (m, 2H), 1.70-1.74 (m, 1H), 1.89-1.93 (m, 2H), 2.38(s, 3H), 2.79-2.90 (m, 2H), 3.85 (t, J=5.9, 1H), 4.29 (t, J=6.9, 1H),6.90 (d, J=6.2, 2H), 7.08-7.29 (m, 6H), 7.35 (t, J=6.2, 2H), 7.44 (d,J=8.1, 2H), 7.73 (d, J=8.1, 2H). LC-MS: 642.8 (M−H)⁻, 99% pure.

Example 201 Preparation ofNα-(4-aminobenzenesulfonyl)-Nα-isobutyl-Nε-[N′α-(4-methylbenzenesulfonyl)-L-phenylalanyl]-L-lysine

A suspension of Nα-(4-aminobenzenesulfonyl)-Nα-isobutyl-L-lysinepotassium salt (190 mg, 0.5 mmol, example 166, step B) in THF (10 mL)was treated with a solution ofNα-(4-methylbenzenesulfonyl)-L-phenylalanine acid chloride (187 mg, 0.5mmol) in dry THF (10 mL). The suspension was stirred for 4 h.Afterwards, water (2 mL) was added resulting in a clear solution. Then,EtOAc (30 mL) was added and the organic phase was washed with 1N HCl.The organic phase was removed. Evaporation of the solvent gave a crudeproduct which was purified by preparative HPLC to yield 15 mg (4.6%) ofthe title compound.

¹H NMR (CDCl₃): δ 0.74 (d, J=6.3, 3H), 0.80 (d, J=6.3, 3H), 1.00-1.11(m, 4H), 1.23-1.25 (m, 1H), 1.70-1.74 (m, 1H), 1.89-1.93 (m, 1H), 2.32(s, 3H), 2.65 (m, 2H), 2.84-2.95 (ABX, J=10.1, 7.1, 2H), 3.88 (t, J=6.0,1H), 4.11 (t, J=6.9, 1H), 6.84 (d, J=6.6, 1H), 7.02-7.21 (m, 7H), 7.24(d, J=8.0, 1H), 7.73 (d, J=8.1, 2H). LC-MS: 657.9 (M−H)⁻, 99% pure.

TABLE 1 Anti-protease activity of Ne-amino acid substituted L-lysinederivatives and analogs of formula I.

D, L, DL Ex. No. Cx R₁ R₂ R₃ R₄ R₅ Y n Ki (nM) R, S, RS 1 COOH i-C₄H₉4-CH₃C₆H₄SO₂ H 4-CH₃C₆H₄SO₂ C₆H₅CH₂ O 4 1.1 L, L 2 COOH i-C₄H₉4-CH₃C₆H₄SO₂ H 4-CH₃C₆H₄SO₂ Indole-3-CH₂ O 4 0.513 L, L 3 COOH i-C₄H₉4-CH₃C₆H₄SO₂ H 4-AcNHC₆H₄SO₂ C₆H₅CH₂ O 4 0.977 L, L 4 COOH i-C₄H₉4-CH₃C₆H₄SO₂ H C₆H₅SO₂ Indole-3-CH₂ O 4 1.0 L, L 5 COOH i-C₄H₉4-NH₂C₆H₄SO₂ H 4-NH₂C₆H₄SO₂ Indole-3-CH₂ O 4 0.459 L, L 6 COOH i-C₄H₉4-CH₃C₆H₄SO₂ H 4-NO₂C₆H₄SO₂ Indole-3-CH₂ O 4 7.3 L, L 7 COOH i-C₄H₉4-CH₃C₆H₄SO₂ H 4-CH₃C₆H₄SO₂ C₆H₅CH₂ O 4 120 D, L 8 COOH i-C₄H₉4-CH₃C₆H₄SO₂ H C₆H₅SO₂ C₆H₅CH₂ O 4 1.9 L, L 9 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-CIC₆H₄SO₂ C₆H₅CH₂ O 4 6.8 L, L 10 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-NO₂C₆H₄SO₂ C₆H₅CH₂ O 4 18 L, L 11 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-CH₃C₆H₄SO₂ 4-HOC₆H₄CH₂ O 4 N/A L, L 12 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-NH₂C₆H₄SO₂ Indole-3-CH₂ O 4 0.311 L, L 13 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-NO₂C₆H₄SO₂ CH₃ O 4 N/A L, L 14 COOH i-C₄H₉ 4-CH₃C₅H₄SO₂ H C₆H₅SO₂CH₃CH₂CH₂ O 4 185 L, L 15 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H C₆H₅SO₂CH₃CH₂CH₂CH₂ O 4 34 L, L 16 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H 4-NO₂C₆H₄SO₂i-C₄H₉ O 4 34 L, L 17 See Table 2 18 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-FC₆H₄SO₂ C₆H₅CH₂ O 4 8.3 L, L 19 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H2-Naphthyl-SO₂ C₆H₅CH₂ O 4 11 L, L 20 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-BrC₅H₄SO₂ C₆H₅CH₂ O 4 19 L, L 21 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-CH₃C₆H₄SO₂ H O 4 21 L 22 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H C₆H₅SO₂ i-C₄H₉ O 429 L, L 23 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H 4-CF₃C₆H₄SO₂ C₆H₅CH₂ O 4 12 L, L24 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H 2-Thiophene-SO₂ C₆H₅CH₂ O 4 0.506 L, L 25COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H C₆H₅SO₂ H₂NC(O)CH₂ O 4 5.3 L, L 26 COOHi-C₄H₉ 4-CH₃C₆H₄SO₂ H 4-CH₃C₆H₄SO₂ 4-NO₂C₆H₄CH₂ O 4 2.6 L, L 27 COOHi-C₄H₉ 4-CH₃C₅H₄SO₂ H 4-CH₃C₆H₄SO₂ C₆H₅ O 4 11 L, L 28 COOH i-C₄H₉4-NO₂C₆H₄SO₂ H 4-AcNHC₆H₄SO₂ C₆H₅CH₂ O 4 9.3 L, L 29 COOH i-C₄H₉4-NO₂C₆H₄SO₂ H 2-Thiophene-SO₂ C₆H₅CH₂ O 4 0.506 L, L 30 COOH i-C₄H₉4-CH₃C₆H₄SO₂ H Ac C₆H₅CH₂ O 4 2.6 L, L 31 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ HC₆H₅CH2O₂C C₆H₅CH₂ O 4 2.3 L, L 32 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-CH₃C₆H₄SO₂ HOCH₂ O 4 93 L, L 33 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-CH₃C₆H₄SO₂ C₆H₁₁CH₂ O 4 3.2 L, L 34 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-CH₃C₆H₄SO₂ H₂NC(O)CH₂CH₂ O 4 95 L, L 35 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-CH₃C₆H₄SO₂ 2-Thiophene-CH₂ O 4 21 L, L 36 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ HFmoc HOCH₂ O 4 162 L, L 37 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H Fmoc C₆H₁₁CH₂ O 428 L, L 38 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H 4-CH₃C₆H₄SO₂ HO₂CCH₂CH₂ O 4 220 L,L 39 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H 4-CH₃C₆H₄SO₂ H₂NCH₂CH₂CH₂CH₂ O 4 >300 L,L 40 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H 4-CH₃C₆H₄SO₂ C₆H₅CH₂OCH₂ O 4 8.0 L, L 41COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H 4-CH₃C₆H₄SO₂ HO₂CCH₂ O 4 172 L, L 42 COOHi-C₄H₉ 4-CH₃C₆H₄SO₂ H 4-CH₃C₆H₄SO₂ 2-Benzothiophene-CH₂ O 4 256 L, L 43COOH i-C₄H₉ 4-NH₂C₆H₄SO₂ H 4-CH₃C₆H₄SO₂ Indole-3-CH₂ O 4 0.56 L, L 44COOH i-C₄H₉ 4-NH₂C₆H₄SO₂ H C₆H₅SO₂ C₆H₅CH₂ O 4 3.6 L, L 45COOCH₂CH(OH)CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H 4-CH₃C₆H₄SO₂ Indole-3-CH₂ O4 >300 L, L 46 COOH i-C₄H₉ 4-NH₂C₆H₄SO₂ H 2-Thiophene-SO₂ C₆H₅CH₂ O 40.477 L, L 47 COOH i-C₄H₉ 4-NH₂C₆H₄SO₂ H 4-CH₃C₆H₄SO₂ H₂NC(O)CH₂ O 4 54L, L 48 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H C₆H₅CH₂O₂C H₂NC(O)CH₂ O 4 114 L, L 49CONHNH₂ i-C₄H₉ 4-NH₂C₆H₄SO₂ H 2-Thiophene-SO₂ C₆H₅CH₂ O 4 17 L, L 50COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H C₆H₅CO C₆H₅CH₂ O 4 2.0 L, L 51 COOH i-C₄H₉4-CH₃C₆H₄SO₂ H 4-Morpholine-CO C₆H₅CH₂ O 4 0.9 L, L 52 COOH i-C₄H₉4-CH₃C₆H₄SO₂ H t-butyl-CO C₆H₅CH₂ O 4 0.545 L, L 53 COOH i-C₄H₉4-CH₃C₆H₄SO₂ H 2-Thiophene-SO₂ lndole-3-CH₂ O 4 13 L, L 54 COOH i-C₄H₉2-Thiophene-SO₂ H 4-CH₃C₆H₄SO₂ C₆H₅CH₂ O 4 17 L, L 55 COOH i-C₄H₉4-CH₃C₆H₄SO₂ i-C₄H₉ 4-CH₃C₆H₄SO₂ H O 4 14 L 56 COOH i-C₄H₉ 4-NH₂C₆H₄SO₂H 4-AcNHC₆H₄SO₂ C₆H₅CH₂ O 4 1.9 L, L 57 COOCH₃ i-C₄H₉ 4-CH₃C₆H₄SO₂ H2-Thiophene-SO₂ C₆H₅CH₂ O 4 >300 L, L 58 CONH₂ i-C₄H₉ 4-CH₃C₆H₄SO₂ H2-Thiophene-SO₂ C₆H₅CH₂ O 4 5.2 L, L 59 CONHOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H2-Thiophene-SO₂ C₆H₅CH₂ O 4 13 L, L 60 CONHCH₂CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂H 2-Thiophene-SO₂ C₆H₅CH₂ O 4 >300 L, L 61 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-CH₃C₆H₄SO₂ H₂NC(O)CH₂ O 4 296 L, L 62 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ Ht-butyl-CO H₂NC(O)CH₂ O 4 212 L, L 63 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H C₆H₅COH₂NC(O)CH₂ O 4 193 L, L 64 CONHNH₂ i-C₄H₉ 4-CH₃C₆H₄SO₂ H 2-Thiophene-SO₂C₆H₅CH₂ O 4 >300 L, L 65 See Table 2 66 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-CH₃C₆H₄SO₂ C₆H₅CH₂ O 4 >300 D, D 67 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-CH₃C₆H₄SO₂ C₆H₅CH₂ O 4 >300 L, D 68 COOCH₃ i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-CH₃C₆H₄SO₂ C₆H₅CH₂ O 4 >300 L, L 69 COOCH₃ i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-CH₃C₆H₄SO₂ Indole-3-CH₂ O 4 >300 L, L 70 COOCH₃ i-C₄H₉ 4-CH₃C₆H₄SO₂ HC₆H₅CO C₆H₅CH₂ O 4 >300 L, L 71 COOCH₃ i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-CH₃C₆H₄SO₂ C₆H₅CH₂ S 4 >300 L, L 72 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-CH₃C₆H₄SO₂ C₆H₅CH₂ S 4 33 L, L 73 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-CH₃C₆H₄SO₂ Indole-3-CH₂ S 4 14 L, L 74 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ HC₆H₅SO₂ Indole-3-CH₂ S 4 4.8 L, L 75 COOCH₃ i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-CH₃C₆H₄SO₂ Indole-3-CH₂ S 4 >300 L, L 76 COOCH₃ i-C₄H₉ 4-CH₃C₆H₄SO₂ HC₆H₅CS C₆H₅CH₂ S 4 >300 L, L 77 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H C₆H₅CSC₆H₅CH₂ S 4 212 L, L 78 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H Boc C₆H₄CH₂ O 4 2.5L, L 79 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H H C₆H₅CH₂ O 4 1.5 L, L 80 COOH i-C₄H₉4-CH₃C₆H₄SO₂ H Carbotetrahydro-3-furanyloxy C₆H₅CH₂ O 4 1.4 L, L 81 COOHi-C₄H₉ 4-CH₃C₆H₄SO₂ CH₃ Boc C₆H₄CH₂ O 4 1.4 L, L 82 COOH i-C₄H₉4-CH₃C₆H₄SO₂ H Boc CH₃SCH₂CH₂ O 4 65 L, L 83 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ HBoc 4-CH₃C₆H₄CH₂SCH₂ O 4 62 L, L 84 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H BocBnOCH(CH₃) O 4 163 L, L 85 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H BocNτ-Bn-imidazole-4-CH₂ O 4 124 L, L 86 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H BocIndole-3-CH₂ O 4 1.6 L, L 87 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H Boc4-BnO-C₆H₄CH₂ O 4 49 L, L 88 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H CH₃ C₆H₅CH₂ O 42.1 L, L 89 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H H CH₃SCH₂CH₂ O 4 33 L, L 90 COOHi-C₄H₉ 4-CH₃C₆H₄SO₂ H H 4-CH₃C₆H₄CH₂SCH₂ O 4 29 L, L 91 COOH i-C₄H₉4-CH₃C₆H₄SO₂ H H BnOCH(CH₃) O 4 10 L, L 92 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H HNτ-Bn-imidazole-4-CH₂ O 4 >300 L, L 93 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H HIndole-3-CH₂ O 4 2.1 L, L 94 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H H 4-BnO—C₆H₄CH₂O 4 177 L, L 95 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H C₆H₅CO C₆H₅CH₂ O 4 54 S, S96 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H 4-CH₃C₆H₄SO₂ C₆H₅CH₂ O 4 16 S, S 97 CH₂OHi-C₄H₉ 4-CH₃C₆H₄SO₂ H 4-CH₃C₆H₄SO₂ Indole-3-CH₂ O 4 3.6 S, S 98 COOHi-C₄H₉ 4-CH₃C₆H₄SO₂ H 4-CH₃C₆H₄SO₂ C₆H₅CH₂ NC—N 4 5.1 L, L 99 COOHi-C₄H₉ 4-CH₃C₆H₄SO₂ H Ac Indole-3-CH₂ O 4 0.438 L, L 100 COOH i-C₄H₉4-CH₃C₆H₄SO₂ H t-butyl-CO Indole-3-CH₂ O 4 0.388 L, L 101 COOH i-C₄H₉4-CH₃C₆H₄SO₂ H CF₃CO C₆H₅CH₂ O 4 2.9 L, L 102 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂H 4-CH₃C₆H₄SO₂ Indole-3-CH₂ S 4 51 S, S 103 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-CH₃C₆H₄SO₂ Indole-3-CH₂ NC—N 4 1.1 L, L 104 COOCH₂CH(OH)CH₂OH i-C₄H₉4-CH₃C₆H₄SO₂ H C₆H₅SO₂ Indole-3-CH₂ O 4 >300 L, L 105 COOH i-C₄H₉4-CH₃C₆H₄SO₂ H C₆H₅ H O 4 7.7 L 106 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H 3-C₅H₄N HO 4 >300 L 107 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H 2,3-(CH₃O)₂Bn H O 4 52 L 108COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H 2-C₅H₄N H O 4 >300 L 109 COOH i-C₄H₉4-CH₃C₆H₄SO₂ H C₆H₅CH₂ H O 4 >300 L 110 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ HC₅H₁₀N—CH₂CH₂ H O 4 >300 L 111 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H OC₄H₆N—CH₂CH₂H O 4 >300 L 112 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H 4-C₅H₄N H O 4 >300 L 113COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H 3-quinolyl H O 4 12 L 114 COOH i-C₄H₉4-CH₃C₆H₄SO₂ C₆H₅ C₆H₅CH₂ H O 4 0.937 L 115 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ CH₃C₆H₅ H O 4 12 L 116 CHO i-C₄H₉ 4-CH₃C₆H₄SO₂ H 4-CH₃C₆H₄SO₂ Indole-3-CH₂O 4 >300 L, L 117 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H t-butyl-CO Indole-3-CH₂ O4 0.428 S, S 118 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H Ac C₆H₅CH₂ O 4 13 S, S 119CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H t-butyl-CO C₆H₅CH₂ O 4 3.9 S, S 120 CH₂OHi-C₄H₉ 4-CH₃C₆H₄SO₂ H 4-Morpholine-CO C₆H₅CH₂ O 4 2.4 S, S 121 CH₂OHi-C₄H₉ 4-CH₃C₆H₄SO₂ H 4-AcNHC₆H₄SO₂ C₆H₅CH₂ O 4 23 S, S 122 CH₂OH i-C₄H₉4-CH₃C₆H₄SO₂ H 2-Thiophene-SO₂ C₆H₅CH₂ O 4 8.2 S, S 123 CH₂OH i-C₄H₉4-CH₃C₆H₄SO₂ H C₆H₅SO₂ C₆H₅CH₂ O 4 14 S, S 124 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂H 4-CH₃C₆H₄SO₂ 4-NO₂C₆H₄CH₂ O 4 12 S, S 125 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-FC₆H₄SO₂ C₆H₅CH₂ O 4 39 S, S 126 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-CH₃OC₆H₄SO₂ C₆H₅CH₂ O 4 19 S, S 127 CH₂OH i-C₄H₉ 4-CH₃C₅H₄SO₂ H4-CF₃C₆H₄SO₂ C₆H₅CH₂ O 4 104 S, S 128 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ HC₆H₅SO₂ Indole-3-CH₂ O 4 0.95 S, S 129 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H2-Thiophene-SO₂ Indole-3-CH₂ O 4 1.0 S, S 130 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂H 4-NO₂C₆H₄SO₂ Indole-3-CH₂ O 4 27 S, S 131 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ HAc Indole-3-CH₂ O 4 3.0 S, S 132 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H4-CH₃OC₆H₄CH₂CO C₆H₅CH₂ O 4 0.257 L, L 133 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ HC₆H₅CH₂CH₂CO Indole-3-CH₂ O 4 0.606 L, L 134 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ HBoc C₅H₄N-2-CH₂ O 4 30 S, S 135 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H BocC₅H₄N-3-CH₂ O 4 144 S, S 136 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H Boc C₅H₄N-4-CH₂O 4 110 S, S 137 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H Boc Thiazole-4-CH₂ O 4 96S, S 138 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H Boc 2-FC₆H₅CH₂ O 4 10 S, S 139CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H Boc 3-FC₆H₅CH₂ O 4 4.1 S, S 140 CH₂OH i-C₄H₉4-CH₃C₆H₄SO₂ H Boc 4-FC₆H₅CH₂ O 4 4.2 S, S 141 See Table 2 142 CH₂OHi-C₄H₉ 4-CH₃C₅H₄SO₂ H Boc 4-t-Butyl-C₆H₅CH₂ O 4 93 S, S 143 CH₂OH i-C₄H₉4-CH₃C₆H₄SO₂ H Boc C₆F₅CH₂ O 4 >300 S, S 144 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ HBoc 4-FmocNHCH₂—C₆H₅CH₂ O 4 >300 S, S 145 CH₂OH i-C₄H₉ 4-NH₂C₆H₄SO₂ H4-CH₃C₆H₄SO₂ Indole-3-CH₂ O 4 1.9 S, S 146 CH₂OH i-C₄H₉ 4-CH₃OC₆H₄SO₂ HH TFA Indole-3-CH₂ O 4 15 S, S 147 CH₂OH i-C₄H₉ 4-CH₃OC₆H₄SO₂ H BocIndole-3-CH₂ O 4 16 S, S 148 CH₂OH i-C₄H₉ 4-CH₃OC₆H₄SO₂ H t-butyl-COIndole-3-CH₂ O 4 1.9 S, S 149 CH₂OH i-C₄H₉ 4-CH₃OC₆H₄SO₂ H 4-CH₃C₆H₄SO₂Indole-3-CH₂ O 4 7.5 S, S 150 COOCH₃ Cyclopentyl-CH₂ 4-CH₃OC₆H₄SO₂ H BocIndole-3-CH₂ O 4 >300 L, L 151 COOH Cyclopentyl-CH₂ 4-CH₃OC₆H₄SO₂ H BocIndole-3-CH₂ O 4 5.3 L, L 152 CH₂OH Cyclopentyl-CH₂ 4-CH₃OC₆H₄SO₂ H HTFA Indole-3-CH₂ O 4 12 S, S 153 CH₂OH Cyclopentyl-CH₂ 4-CH₃OC₆H₄SO₂ HBoc Indole-3-CH₂ O 4 24 S, S 154 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H 4-CH₃C₆H₄SO₂C₆H₅CH₂ O 3 >300 L, L 155 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ C₆H₅CH₂ C₆H₅CH₂ H O 4N/A L 156 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ C₆H₅CH₂ C₆H₅ H O 4 7.6 S 157 COOHi-C₄H₉ 4-CH₃C₆H₄SO₂ H C₆H₁₁ H O 4 N/A L 158 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂C₆H₅CH₂ C₆H₅CO H O 4 95 S 159 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ C₆H₅CH₂4-CH₃C₆H₄SO₂ H O 4 101 S 160 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ C₆H₅ C₆H₅CH₂CH₂ HO 4 52 S 161 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H 4-NH₂C₆H₄SO₂ Indole-3-CH₂ O 41.4 S, S 162 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H 4-CH₃OC₆H₄CH₂CO C₆H₅CH₂ O 4 1.2S, S 163 COOH i-C₄H₉ 4-NH₂C₅H₄SO₂ H 4-CH₃OC₆H₄CH₂CO C₆H₅CH₂ O 4 0.270 L,L 164 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ H 4-CH₃OC₆H₄CH₂CO Indole-3-CH₂ O 4 0.320L, L 165 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H 4-CH₃OC₆H₄CH₂CO Indole-3-CH₂ O 41.1 S, S 166 CH₂OH i-C₄H₉ 4-NH₂C₆H₄SO₂ H 2-Thiophene-SO₂ Indole-3-CH₂ O4 1.0 S, S 167 CH₂OH i-C₄H₉ 4-NH₂C₆H₄SO₂ H C₆H₅SO₂ Indole-3-CH₂ O 4 1.5S, S 168 CH₂OH i-C₄H₉ 4-NH₂C₆H₄SO₂ H Ac Indole-3-CH₂ O 4 3.8 S, S 169CH₂OH i-C₄H₉ 4-NH₂C₆H₄SO₂ H t-butyl-CO Indole-3-CH₂ O 4 1.2 S, S 170CH₂OH i-C₄H₉ 4-NH₂C₆H₄SO₂ H 4-Morpholine-CO Indole-3-CH₂ O 4 1.1 S, S171 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H 4-Morpholine-CO Indole-3-CH₂ O 4 7.9 S,S 172 CH₂OH i-C₄H₉ 4-CH₃OC₆H₄SO₂ H Ac Indole-3-CH₂ O 4 >300 S, S 173CH₂OH Cydopentyl-CH₂ 4-CH₃OC₆H₄SO₂ H Ac Indole-3-CH₂ O 4 11 S, S 174COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ C₆H₅ 4-CH₃OC₆H₄CH₂ H O 4 1.4 L 175 CH₂OH i-C₄H₉4-CH₃C₆H₄SO₂ C₆H₅ 4-CH₃OC₆H₄CH₂ H O 4 6.3 S 176 CH₂OH i-C₄H₉4-NH₂C₆H₄SO₂ C₆H₅ 4-CH₃OC₆H₄CH₂ H O 4 2.4 S 177 CH₂OH i-C₄H₉4-NH₂C₆H₄SO₂ H CH₃O—CO C₆H₅CH₂ O 4 6.6 S, S 178 CH₂OH i-C₄H₉4-NH₂C₆H₄SO₂ H CH₃OCO Indole-3-CH₂ O 4 2.0 S, S 179 CH₂OH i-C₄H₉4-NH₂C₆H₄SO₂ H 4-NO₂C₆H₄SO₂ Indole-3-CH₂ O 4 32 S, S 180 CH₂OH i-C₄H₉4-NH₂C₆H₄SO₂ H 4-Morphoine-CO C₆H₅CH₂ O 4 2.4 S, S 181 CH₂OH i-C₄H₉4-NH₂C₆H₄SO₂ i-C₄H₉ C₆H₅ H O 4 16 S 182 CH₂OH i-C₄H₉ 4-NH₂C₆H₄SO₂C₆H₅CH₂ C₆H₅ H O 4 2.4 S 183 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H H TFAIndole-3-CH₂ O 4 4.4 S, S 184 CH₂OH i-C₄H₉ 4-NH₂C₆H₄SO₂ H i-Pr-COIndole-3-CH₂ O 4 1.6 S, S 185 CH₂OH i-C₄H₉ 4-NH₂C₆H₄SO₂ H t-butyl-CH₂COIndole-3-CH₂ O 4 3.1 S, S 186 CH₂OH i-C₄H₉ 4-NH₂C₆H₄SO₂ H C₆H₅COIndole-3-CH₂ O 4 1.7 S, S 187 CH₂OH i-C₄H₉ 4-NH₂C₆H₄SO₂ H 4-NH₂C₆H₄SO₂Indole-3-CH₂ O 4 0.200 S, S 188 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H C₆H₅SO₂NC-CH₂ O 4 79 S, S 189 CONH₂ i-C₄H₉ 4-CH₃C₆H₄SO₂ H C₆H₅SO₂ Indole-3-CH₂O 4 1.9 L, L 190 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H CH₃O—CO Indole-3-CH₂ O 42.2 S, S 191 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H C₆H₅CO Indole-3-CH₂ O 4 3.8 S,S 192 CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ H i-butylO—CO Indole-3-CH₂ O 4 4.5 S 193CH₂OH i-C₄H₉ 4-NH₂C₆H₄SO₂ H i-butylO—CO Indole-3-CH₂ O 4 4.1 S 194 CH₂OHi-C₄H₉ 4-NH₂C₆H₄SO₂ H 4-NO₂C₆H₄SO₂ C₆H₅CH₂ O 4 85 S 195 CH₂OH i-C₄H₉4-NH₂C₆H₄SO₂ H 4-NH₂C₆H₄SO₂ C₆H₅CH₂ O 4 20 S 196 COOH i-C₄H₉4-CH₃C₆H₄SO₂ H 4-CH₃OC₆H₄NHCO C₆H₅CH₂ O 4 0.500 L 197 COOH i-C₄H₉4-CH₃C₆H₄SO₂ H Pyrrolidine-CO C₆H₅CH₂ O 4 1.7 L 198 COOH i-C₄H₉4-CH₃C₆H₄SO₂ H CH₃NHCO C₆H₅CH₂ O 4 1.1 L 199 COOH i-C₄H₉ 4-CH₃C₆H₄SO₂ HHOCH₂CH₂NHCO C₆H₅CH₂ O 4 2.2 L 200 COOH i-C₄H₉ C₆H₄SO₂ H 4-CH₃C₆H₄SO₂C₆H₅CH₂ O 4 31 L 201 COOH i-C₄H₉ 4-NH₂C₆H₄SO₂ H 4-CH₃C₆H₄SO₂ C₆H₅CH₂ O 44.1 L

TABLE 2 Anti-protease activity of amino acid derivatives of formula II.

D, L, DL Ex. NO. Cx R₁ R₂ R₄ R₃ Y n Ki (nM) R, S, RS 17 COOH i-C₄H₉4-CH₃C₆H₄SO₂ C₆H₅SO₂ N′αCH₂CH(OH)CH₂ O 4 55 L, L 65 COOH i-C₄H₉4-CH₃C₆H₄SO₂ 4-CH₃C₆H₄SO₂

O 4 24 L, L 141  CH₂OH i-C₄H₉ 4-CH₃C₆H₄SO₂ Boc indole-(2-CH₂N′α)-3-CH₂—O 4 38 S, S

We claim:
 1. A compound selected from the group consisting of a compoundof formula (1)

wherein n is 3 or 4 wherein each m is independently 0 or 1 wherein eacho is independently 0 or 1 wherein R₁ is selected from the groupconsisting of a straight or branched alkyl group of 1 to 6 carbon atoms,and a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkylpart thereof and 1 to 3 carbon atoms in the alkyl part thereof, whereinR₃ is selected from the group consisting of H, a straight or branchedalkyl group of 1 to 6 carbon atoms, a phenyl and a benzyl group whereinR_(12,) R_(12A,) R₁₃ and R_(13A) are independently selected from thegroup consisting of H, a straight or branched alkyl group of 1 to 4carbon atoms, F, Cl, Br, I, —CF₃, —NO₂, —NR₁₀R₁₁, —NHCOR₁₀, —OR₁₀,—OCH₂C₆H₅, —SR₁₀, —COOR₁₀, —COR₁₀ and —CH₂OH, and wherein R₁₀ and R₁₁are independently selected from the group consisting of H, and astraight or branched alkyl group of 1 to 4 carbon atoms, and when one ormore of R_(12,) R_(12A,) R₁₃ and R_(13A) comprises an amino group, or apharmaceutically acceptable ammonium salt thereof.
 2. A compound offormula (1A)

wherein n is 3 or 4 wherein each m is independently 0 or 1 wherein eacho is independently 0 or 1 wherein R₁ is selected from the groupconsisting of a straight or branched alkyl group of 1 to 6 carbon atoms,and a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkylpart thereof and 1 to 3 carbon atoms in the alkyl part thereof, whereinR₃ is selected from the group consisting of H, a straight or branchedalkyl group of 1 to 6 carbon atoms, a phenyl and a benzyl group whereinR_(12,) R_(12A,) R₁₃ and R_(13A) are independently selected from thegroup consisting of H, a straight or branched alkyl group of 1 to 4carbon atoms, F, Cl, Br, I, —CF₃, —NO₂, —NR₁₀R₁₁, —NHCOR₁₀, —OR₁₀,—OCH₂C₆H₅, —SR₁₀, —COOR₁₀, —COR₁₀ and —CH₂OH, and wherein R₁₀ and R₁₁are independently selected from the group consisting of H, and astraight or branched alkyl group of 1 to 4 carbon atoms, and when one ormore of R_(12,) R_(12A,) R₁₃ and R_(13A) comprises an amino group, or apharmaceutically acceptable ammonium salt thereof.
 3. A compound asdefined in claim 2 wherein n is
 4. 4. A compound as defined in claim 3wherein R₁ is iso-butyl.
 5. A compound of formula (1A) as defined inclaim 2 wherein each o is 0, each m is 1, R₁ is selected from the groupconsisting of iso-butyl and cyclopentyl-CH₂— and R₁₂ and R_(12A) areindependently selected from the group consisting of H, —CH₃, —NH₂, —NO₂and —OCH₃.
 6. A compound of formula (1A) as defined in claim 5 whereinR₁ is iso-butyl.
 7. A compound of formula


8. A compound of formula

or a pharmaceutically acceptable ammonium salt thereof.
 9. A compound offormula

or a pharmaceutically acceptable ammonium salt thereof.
 10. A compoundof formula

or a pharmaceutically acceptable ammonium salt thereof.
 11. A compoundof formula


12. A compound of formula

or a pharmaceutically acceptable ammonium salt thereof.
 13. Apharmaceutical composition comprising a pharmaceutically acceptablecarrier and a pharmaceutically effective amount of at least one compoundas defined in claim
 2. 14. A compound selected from the group consistingof a compound of formula (1a)

wherein n is 3 or 4 wherein each m is independently 0 or 1 wherein eacho is independently 0 or 1 wherein R₁ is selected from the groupconsisting of a straight or branched alkyl group of 1 to 6 carbon atoms,and a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkylpart thereof and 1 to 3 carbon atoms in the alkyl part thereof, whereinR_(2A) and R_(4A) are independently selected from the group consistingof H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl,Br, I, —CF₃, —NO₂, —NR₁₀R₁₁, —NHCOR₁₀, —OR₁₀, —OCH₂C₆H₅, —SR₁₀, —COOR₁₀,—COR₁₀ and —CH₂OH, and wherein R₁₀ and R₁₁ are independently selectedfrom the group consisting of H, and a straight or branched alkyl groupof 1 to 4 carbon atoms, and when one of R_(2A,) and R_(4A) or bothcomprises an amino group, or a pharmaceutically acceptable ammonium saltthereof.
 15. A compound of formula (1a) as defined in claim 14 whereineach o is 0, each m is 1, R₁ is selected from the group consisting ofiso-butyl and cyclopentyl-CH₂— and R_(2A,) and R_(4A) are independentlyselected from the group consisting of H, —CH₃, —NH₂, —NO₂ and —OCH₃. 16.A compound of formula (1a) as defined in claim 15 wherein R₁ isiso-butyl.
 17. A pharmaceutical composition comprising apharmaceutically acceptable carrier and a pharmaceutically effectiveamount of at least one compound as defined in claim 14.